Apolipoprotein A-I agonists and their use to treat dyslipidemic disorders

ABSTRACT

The present invention provides peptides and peptide analogues that mimic the structural and pharmacological properties of human ApoA-I. The peptides and peptide analogues are useful to treat a variety of disorders associated with dyslipidemia.

1. INTRODUCTION

The invention relates to apolipoprotein A-I (ApoA-I) agonistcompositions for treating disorders associated with dyslipoproteinemia,including hypercholesterolemia, cardiovascular disease, atherosclerosis,restenosis, and other disorders such as septic shock.

2. BACKGROUND OF THE INVENTION

Circulating cholesterol is carried by plasma lipoproteins--particles ofcomplex lipid and protein composition that transport lipids in theblood. Low density lipoproteins (LDL), and high density lipoproteins(HDL) are the major cholesterol carriers. LDL are believed to beresponsible for the delivery of cholesterol from the liver (where it issynthesized or obtained from dietary sources) to extrahepatic tissues inthe body. The term "reverse cholesterol transport" describes thetransport of cholesterol from extrahepatic tissues to the liver where itis catabolized and eliminated. It is believed that plasma HDL particlesplay a major role in the reverse transport process, acting as scavengersof tissue cholesterol.

The evidence linking elevated serum cholesterol to coronary heartdisease is overwhelming. For example, atherosclerosis is a slowlyprogressive disease characterized by the accumulation of cholesterolwithin the arterial wall. Compelling evidence supports the concept thatlipids deposited in atherosclerotic lesions are derived primarily fromplasma LDL; thus, LDLs have popularly become known as the "bad"cholesterol. In contrast, HDL serum levels correlate inversely withcoronary heart disease--indeed, high serum levels of HDL are regarded asa negative risk factor. It is hypothesized that high levels of plasmaHDL are not only protective against coronary artery disease, but mayactually induce regression of atherosclerotic plaques (e.g., see Badimonet al., 1992, Circulation 86 (Suppl. III):86-94). Thus, HDL havepopularly become known as the "good" cholesterol.

2.1. Cholesterol Transport

The fat-transport system can be divided into two pathways: an exogenousone for cholesterol and triglycerides absorbed from the intestine, andan endogenous one for cholesterol and triglycerides entering thebloodstream from the liver and other non-hepatic tissue.

In the exogenous pathway, dietary fats are packaged into lipoproteinparticles called chylomicrons which enter the bloodstream and delivertheir triglycerides to adipose tissue (for storage) and to muscle (foroxidation to supply energy). The remnant of the chylomicron, containingcholesteryl esters, is removed from the circulation by a specificreceptor found only on liver cells. This cholesterol then becomesavailable again for cellular metabolism or for recycling to extrahepatictissues as plasma lipoproteins.

In the endogenous pathway, the liver secretes a large, very-low-densitylipoprotein particle (VLDL) into the bloodstream. The core of VLDLsconsists mostly of triglycerides synthesized in the liver, with asmaller amount of cholesteryl esters (either synthesized in the liver orrecycled from chylomicrons). Two predominant proteins are displayed onthe surface of VLDLs, apoprotein B-100 and apoprotein E. When a VLDLreaches the capillaries of adipose tissue or of muscle, itstriglycerides are extracted resulting in a new kind of particle,decreased in size and enriched in cholesteryl esters but retaining itstwo apoproteins, called intermediate-density lipoprotein (IDL).

In human beings, about half of the IDL particles are removed from thecirculation quickly (within two to six hours of their formation),because they bind tightly to liver cells which extract their cholesterolto make new VLDL and bile acids. The IDL particles which are not takenup by the liver remain in the circulation longer. In time, theapoprotein E dissociates from the circulating particles, converting themto LDL having apoprotein B-100 as their sole protein.

Primarily, the liver takes up and degrades most of the cholesterol tobile acids, which are the end products of cholesterol metabolism. Theuptake of cholesterol containing particles is mediated by LDL receptors,which are present in high concentrations on hepatocytes. The LDLreceptor binds both apoprotein E and apoprotein B-100, and isresponsible for binding and removing both IDLs and LDLs from thecirculation. However, the affinity of apoprotein E for the LDL receptoris greater than that of apoprotein B-100. As a result, the LDL particleshave a much longer circulating life span than IDL particles--LDLscirculate for an average of two and a half days before binding to theLDL receptors in the liver and other tissues. High serum levels of LDL(the "bad" cholesterol) are positively associated with coronary heartdisease. For example, in atherosclerosis, cholesterol derived fromcirculating LDLs accumulates in the walls of arteries leading to theformation of bulky plaques that inhibit the flow of blood until a cloteventually forms, obstructing the artery causing a heart attack orstroke.

Ultimately, the amount of intracellular cholesterol liberated from theLDLs controls cellular cholesterol metabolism. The accumulation ofcellular cholesterol derived from VLDLs and LDLs controls threeprocesses: first, it reduces cellular cholesterol synthesis by turningoff the synthesis of HMGCoA reductase--a key enzyme in the cholesterolbiosynthetic pathway. Second, the incoming LDL-derived cholesterolpromotes storage of cholesterol by activating ACAT--the cellular enzymewhich converts cholesterol into cholesteryl esters that are deposited instorage droplets. Third, the accumulation of cholesterol within the celldrives a feedback mechanism that inhibits cellular synthesis of new LDLreceptors. Cells, therefore, adjust their complement of LDL receptors sothat enough cholesterol is brought in to meet their metabolic needs,without overloading. (For a review, see Brown & Goldstein, In, ThePharmacological Basis Of Therapeutics, 8th Ed., Goodman & Gilman,Pergamon Press, New York, 1990, Ch. 36, pp. 874-896).

2.2. Reverse Cholesterol Transport

In sum, peripheral (non-hepatic) cells obtain their cholesterol from acombination of local synthesis and the uptake of preformed sterol fromVLDLs and LDLs. In contrast, reverse cholesterol transport (RCT) is thepathway by which peripheral cell cholesterol can be returned to theliver for recycling to extrahepatic tissues, or excretion into theintestine in bile, either in modified or in oxidized form as bile acids.The RCT pathway represents the only means of eliminating cholesterolfrom most extrahepatic tissues, and is crucial to maintenance of thestructure and function of most cells in the body.

The RCT consists mainly of three steps: (a) cholesterol efflux, theinitial removal of cholesterol from various pools of peripheral cells;(b) cholesterol esterification by the action of lecithin:cholesterolacyltransferase (LCAT), preventing a re-entry of effluxed cholesterolinto cells; and (c) uptake/delivery of HDL cholesteryl ester to livercells. The RCT pathway is mediated by HDLs. HDL is a generic term forlipoprotein particles which are characterized by their high density. Themain lipidic constituents of HDL complexes are various phospholipids,cholesterol (ester) and triglycerides. The most prominent apolipoproteincomponents are A-I and A-II which determine the functionalcharacteristics of HDL; furthermore minor amounts of apolipoprotein C-I,C-II, C-III, D, E, J, etc. have been observed. HDL can exist in a widevariety of different sizes and different mixtures of the above-mentionedconstituents depending on the status of remodeling during the metabolicRCT cascade.

The key enzyme involved in the RCT pathway is LCAT. LCAT is producedmainly in the liver and circulates in plasma associated with the HDLfraction. LCAT converts cell derived cholesterol to cholesteryl esterswhich are sequestered in HDL destined for removal. Cholesteryl estertransfer protein (CETP) and phospholipid transfer protein (PLTP)contribute to further remodeling the circulating HDL population. CETPcan move cholesteryl esters made by LCAT to other lipoproteins,particularly ApoB-containing lipoproteins, such as VLDL and LDL. PLTPsupplies lecithin to HDL. HDL triglycerides can be catabolized by theextracellular hepatic triglyceride lipase, and lipoprotein cholesterolis removed by the liver via several mechanisms.

Each HDL particle contains at least one copy (and usually two to fourcopies) of ApoA-I. ApoA-I is synthesized by the liver and smallintestine as preproapolipoprotein which is secreted as a proprotein thatis rapidly cleaved to generate a mature polypeptide having 243 aminoacid residues. ApoA-I consists mainly of 6 to 8 different 22 amino acidrepeats spaced by a linker moiety which is often proline, and in somecases consists of a stretch made up of several residues. ApoA-I formsthree types of stable complexes with lipids: small, lipid-poor complexesreferred to as pre-beta-1 HDL; flattened discoidal particles containingpolar lipids (phospholipid and cholesterol) referred to as pre-beta-2HDL; and spherical particles containing both polar and nonpolar lipids,referred to as spherical or mature HDL (HDL₃ and HDL₂) Most HDL in thecirculating population contain both ApoA-I and ApoA-II (the second majorHDL protein) and are referred to herein as the AI/AII-HDL fraction ofHDL. However, the fraction of HDL containing only ApoA-I (referred toherein as the AI-HDL fraction) appear to be more effective in RCT.Certain epidemiologic studies support the hypothesis that the AI-HDLfraction is anti-atherogenic. (Parra et al., 1992, Arterioscler. Thromb.12:701-707; Decossin et al., 1997, Eur. J. Clin. Invest. 27:299-307).

Although the mechanism for cholesterol transfer from the cell surface(i.e., cholesterol efflux) is unknown, it is believed that thelipid-poor complex, pre-beta-1 HDL is the preferred acceptor forcholesterol transferred from peripheral tissue involved in RCT. (SeeDavidson et al., 1994, J. Biol. Chem. 269:22975-22982; Bielicki et al.,1992, J. Lipid Res. 33:1699-1709; Rothblat et al., 1992, J. Lipid Res.33:1091-1097; and Kawano et al., 1993, Biochemistry 32:5025-5028; Kawanoet al., 1997, Biochemistry 36:9816-9825). During this process ofcholesterol recruitment from the cell surface, pre-beta-1 HDL is rapidlyconverted to pre-beta-2 HDL. PLTP may increase the rate of pre-beta-2disc formation, but data indicating a role for PLTP in RCT is lacking.LCAT reacts preferentially with discoidal and spherical HDL,transferring the 2-acyl group of lecithin or other phospholipids to thefree hydroxyl residue of cholesterol to generate cholesteryl esters(retained in the HDL) and lysolecithin. The LCAT reaction requiresApoA-I as activator; i.e., ApoA-I is the natural cofactor for LCAT. Theconversion of cholesterol to its ester sequestered in the HDL preventsre-entry of cholesterol into the cell, the result being that cholesterylesters are destined for removal. Cholesteryl esters in the mature HDLparticles in the AI-HDL fraction (i.e., containing ApoA-I and noApoA-II) are removed by the liver and processed into bile moreeffectively than those derived from HDL containing both ApoA-I andApoA-II (the AI/AII-HDL fraction). This may be due, in part, to the moreeffective binding of AI-HDL to the hepatocyte membrane. The existence ofan HDL receptor has been hypothesized, and recently a scavengerreceptor, SR-BI, was identified as an HDL receptor (Acton et al., 1996,Science 271:518-520; Xu et al., 1997, J. Lipid Res. 38:1289-1298). TheSR-BI is expressed most abundantly in steroidogenic tissues (e.g., theadrenals), and in the liver (Landshulz et al., 1996, J. Clin. Invest.98:984-995; Rigotti et al., 1996, J. Biol. Chem. 271:33545-33549).

CETP does not appear to play a major role in RCT, and instead isinvolved in the metabolism of VLDL-and LDL-derived lipids. However,changes in CETP activity or its acceptors, VLDL and LDL, play a role in"remodeling" the HDL population. For example, in the absence of CETP,the HDLs become enlarged particles which are not cleared. (For reviewson RCT and HDLs, see Fielding & Fielding, 1995, J. Lipid Res.36:211-228; Barrans et al., 1996, Biochem. Biophys. Acta. 1300:73-85;Hirano et al., 1997, Arterioscler. Thromb. Vasc. Biol. 17(6):1053-1059).

2.3. Current Treatments for Dyslipoproteinemias

A number of treatments are currently available for lowering serumcholesterol and triglycerides (see, e.g., Brown & Goldstein, supra).However, each has its own drawbacks and limitations in terms ofefficacy, side-effects and qualifying patient population.

Bile-acid-binding resins are a class of drugs that interrupt therecycling of bile acids from the intestine to the liver; e.g.,cholestyramine (Questran Light®, Bristol-Myers Squibb), and colestipolhydrochloride (Colestid®, The Upjohn Company). When taken orally, thesepositively-charged resins bind to the negatively charged bile acids inthe intestine. Because the resins cannot be absorbed from the intestine,they are excreted carrying the bile acids with them. The use of suchresins, however, at best only lowers serum cholesterol levels by about20%, and is associated with gastrointestinal side-effects, includingconstipation and certain vitamin deficiencies. Moreover, since theresins bind other drugs, other oral medications must be taken at leastone hour before or four to six hours subsequent to ingestion of theresin; thus, complicating heart patient's drug regimens.

The statins are cholesterol lowering agents that block cholesterolsynthesis by inhibiting HMGCoA reductase--the key enzyme involved in thecholesterol biosynthetic pathway. The statins, e.g., lovastatin(Mevacor®, Merck & Co., Inc.), and pravastatin (Pravachol®,Bristol-Myers Squibb Co.) are sometimes used in combination withbile-acid-binding resins. The statins significantly reduce serumcholesterol and LDL-serum levels, and slow progression of coronaryatherosclerosis. However, serum HDL cholesterol levels are onlymoderately increased. The mechanism of the LDL lowering effect mayinvolve both reduction of VLDL concentration and induction of cellularexpression of LDL-receptor, leading to reduced production and/orincreased catabolism of LDLs. Side effects, including liver and kidneydysfunction are associated with the use of these drugs (Physicians DeskReference, Medical Economics Co., Inc., Montvale, N.J., 1997). Recently,the FDA has approved atorvastatin (an HMGCoA reductase inhibitordeveloped by Parke-Davis) (Warner Lambert) for the market to treat rarebut urgent cases of familial hypercholesterolemia (1995, Scrip20(19):10).

Niacin, or nicotinic acid, is a water soluble vitamin B-complex used asa dietary supplement and antihyperlipidemic agent. Niacin diminishesproduction of VLDL and is effective at lowering LDL. In some cases, itis used in combination with bile-acid binding resins. Niacin canincrease HDL when used at adequate doses, however, its usefulness islimited by serious side effects when used at such high doses.

Fibrates are a class of lipid-lowering drugs used to treat various formsof hyperlipidemia (i.e., elevated serum triglycerides) which may also beassociated with hypercholesterolemia. Fibrates appear to reduce the VLDLfraction and modestly increase HDL--however the effects of these drugson serum cholesterol is variable. In the United States, fibrates havebeen approved for use as antilipidemic drugs, but have not receivedapproval as hypercholesterolemia agents. For example, clofibrate(Atromid-S®, Wyeth-Ayerst Laboratories) is an antilipidemic agent whichacts (via an unknown mechanism) to lower serum triglycerides by reducingthe VLDL fraction. Although serum cholesterol may be reduced in certainpatient subpopulations, the biochemical response to the drug isvariable, and is not always possible to predict which patients willobtain favorable results. Atromid-S® has not been shown to be effectivefor prevention of coronary heart disease. The chemically andpharmacologically related drug, gemfibrozil (Lopid®, Parke-Davis) is alipid regulating agent which moderately decreases serum triglyceridesand VLDL cholesterol, and moderately increases HDL cholesterol--the HDL₂and HDL₃ subfractions as well as both ApoA-I and A-II (i.e., theAI/AII-HDL fraction). However, the lipid response is heterogeneous,especially among different patient populations. Moreover, whileprevention of coronary heart disease was observed in male patientsbetween 40-55 without history or symptoms of existing coronary heartdisease, it is not clear to what extent these findings can beextrapolated to other patient populations (e.g., women, older andyounger males). Indeed, no efficacy was observed in patients withestablished coronary heart disease. Serious side-effects are associatedwith the use of fibrates including toxicity such as malignancy,(especially gastrointestinal cancer), gallbladder disease and anincreased incidence in non-coronary mortality. These drugs are notindicated for the treatment of patients with high LDL or low HDL astheir only lipid abnormality (Physician's Desk Reference, 1997, MedicalEconomics Co., Inc. Montvale, N.J.).

Oral estrogen replacement therapy may be considered for moderatehypercholesterolemia in post-menopausal women. However, increases in HDLmay be accompanied with an increase in triglycerides. Estrogen treatmentis, of course, limited to a specific patient population (postmenopausalwomen) and is associated with serious side effects including inductionof malignant neoplasms, gall bladder disease, thromboembolic disease,hepatic adenoma, elevated blood pressure, glucose intolerance, andhypercalcemia.

Thus, there is a need to develop safer drugs that are efficacious inlowering serum cholesterol, increasing HDL serum levels, preventingcoronary heart disease, and/or treating existing disease, especiallyatherosclerosis.

2.4. ApoA-I as a Target

None of the currently available drugs for lowering cholesterol safelyelevate HDL levels and stimulate RCT--most appear to operate on thecholesterol transport pathway, modulating dietary intake, recycling,synthesis of cholesterol, and the VLDL population.

While it is desirable to find drugs that stimulate cholesterol effluxand removal, several potential targets in the RCT exist--e.g., LCAT, HDLand its various components (ApoA-I, ApoA-II and phospholipids), PLTP,and CETP--and it is not known which target would be most effective atachieving desirable lipoprotein profiles and protective effects.Perturbation of any single component in the RCT pathway ultimatelyaffects the composition of circulating lipoprotein populations, and theefficiency of RCT.

Several lines of evidence based on data obtained in vivo implicate theHDL and its major protein component, ApoA-I, in the prevention ofatherosclerotic lesions, and potentially, the regression ofplaques--making these attractive targets for therapeutic intervention.First, an inverse correlation exists between serum ApoA-I (HDL)concentration and atherogenesis in man (Gordon & Rifkind, 1989, N. Eng.J. Med. 321:1311-1316; Gordon et al., 1989, Circulation 79:8-15).Indeed, specific subpopulations of HDL have been associated with areduced risk for atherosclerosis in humans (Miller, 1987, Amer. Heart113:589-597; Cheung et al., 1991, Lipid Res. 32:383-394); Fruchart &Ailhaud, 1992, Clin. Chem. 38:79).

Second, animal studies support the protective role of ApoA-I (HDL).Treatment of cholesterol fed rabbits with ApoA-I or HDL reduced thedevelopment and progression of plaque (fatty streaks) in cholesterol-fedrabbits. (Koizumi et al., 1988, J. Lipid Res. 29:1405-1415; Badimon etal., 1989, Lab. Invest. 60:455-461; Badimon et al., 1990, J. Clin.Invest. 85:1234-1241). However, the efficacy varied depending upon thesource of HDL (Beitz et al., 1992, Prostaglandins, Leukotrienes andEssential Fatty Acids 47:149-152; Mezdour et al., 1995, Atherosclerosis113:237-246).

Third, direct evidence for the role of ApoA-I was obtained fromexperiments involving transgenic animals. The expression of the humangene for ApoA-I transferred to mice genetically predisposed todiet-induced atherosclerosis protected against the development of aorticlesions (Rubin et al., 1991, Nature 353:265-267). The ApoA-I transgenewas also shown to suppress atherosclerosis in ApoE-deficient mice and inApo(a) transgenic mice (Paszty et al., 1994, J. Clin. Invest.94:899-903; Plump et al., 1994, Proc. Natl. Acad. Sci. U.S.A.91:9607-9611; Liu et al., 1994, J. Lipid Res. 35:2263-2266). Similarresults were observed in transgenic rabbits expressing human ApoA-I(Duverger, 1996, Circulation 94:713-717; Duverger et al., 1996,Arterioscler. Thromb. Vasc. Biol. 16:1424-1429), and in transgenic ratswhere elevated levels of human ApoA-I protected against atherosclerosisand inhibited restenosis following balloon angioplasty (Burkey et al.,1992, Circulation, Supplement I, 86:I-472, Abstract No. 1876; Burkey etal., 1995, J. Lipid Res. 36:1463-1473).

The AI-HDL appear to be more efficient at RCT than the AI/AII-HDLfraction. Studies with mice transgenic for human ApoA-I or Apo-I andApoA-II (AI/AII) showed that the protein composition of HDLsignificantly affects its role--AI-HDL is more anti-atherogenic thanAI/AII-HDL (Schultz et al., 1993, Nature 365:762-764). Parallel studiesinvolving transgenic mice expressing the human LCAT gene demonstratethat moderate increases in LCAT activity significantly changelipoprotein cholesterol levels, and that LCAT has a significantpreference for HDL containing ApoA-I (Francone et al., 1995, J. Clinic.Invest. 96:1440-1448; Berard et al., 1997, Nature Medicine3(7):744-749). While these data support a significant role for ApoA-I inactivating LCAT and stimulating RCT, additional studies demonstrate amore complicated scenario: a major component of HDL that modulatesefflux of cell cholesterol is the phospholipids (Fournier et al., 1996,J. Lipid Res. 37:1704-1711).

In view of the potential role of HDL, i.e., both ApoA-I and itsassociated phospholipid, in the protection against atheroscleroticdisease, human clinical trials utilizing recombinantly produced ApoA-Iwere commenced, discontinued and apparently re-commenced by UCB Belgium(Pharmaprojects, Oct. 27, 1995; IMS R&D Focus, Jun. 30, 1997; DrugStatus Update, 1997, Atherosclerosis 2(6):261-265); see also M. Erikssonat Congress, "The Role of HDL in Disease Prevention," Nov. 7-9, 1996,Fort Worth; Lacko & Miller, 1997, J. Lip. Res. 38:1267-1273; andWO94/13819) and were commenced and discontinued by Bio-Tech(Pharmaprojects, Apr. 7, 1989). Trials were also attempted using ApoA-Ito treat septic shock (Opal, "Reconstituted HDL as a Treatment Strategyfor Sepsis," IBC's 7th International Conference on Sepsis, Apr. 28-30,1997, Washington, D.C.; Gouni et al., 1993, J. Lipid Res. 94:139-146;Levine, WO96/04916). However, there are many pitfalls associated withthe production and use of ApoA-I, making it less than ideal as a drug;e.g., ApoA-I is a large protein that is difficult and expensive toproduce; significant manufacturing and reproducibility problems must beovercome with respect to stability during storage, delivery of an activeproduct and half-life in vivo.

In view of these drawbacks, attempts have been made to prepare peptidesthat mimic ApoA-I. Since the key activities of ApoA-I have beenattributed to the presence of multiple repeats of a unique secondarystructural feature in the protein--a class A amphipathic α-helix(Segrest, 1974, FEBS Lett. 38:247-253), most efforts to design peptideswhich mimic the activity of ApoA-I have focused on designing peptideswhich form class A-type amphipathic α-helices.

Class A-type amphipathic α-helices are unique in that positively chargedamino acid residues are clustered at the hydrophobic-hydrophilicinterface and negatively charged amino acid residues are clustered atthe center of the hydrophilic face. Furthermore, class A α-helicalpeptides have a hydrophobic angle of less than 180° (Segrest et al.,1990, PROTEINS: Structure, Function and Genetics 8:103-117). The initialde novo strategies to design ApoA-I mimics were not based upon theprimary sequences of naturally occurring apolipoproteins, but ratherupon incorporating these unique Class A helix features into thesequences of the peptide analogues, as well as some of the properties ofthe ApoA-I domains (see, e.g., Davidson et al., 1996, Proc. Natl. Acad.Sci. U.S.A. 93:13605-13610; Rogers et al., 1997, Biochemistry36:288-300; Lins et al., 1993, Biochim. Biophys. Acta biomembranes1151:137-142; Ji and Jonas, 1995, J. Biol. Chem. 270:11290-11297; Colletet al., 1997, Journal of Lipid Research, 38:634-644; Sparrow and Gotto,1980, Ann. N.Y. Acad. Sci. 348:187-211; Sparrow and Gotto, 1982, CRCCrit. Rev. Biochem. 13:87-107; Sorci-Thomas et al., 1993, J. Biol. Chem.268:21403-21409; Wang et al., 1996, Biochim. Biophys. Acta 174-184;Minnich et al., 1992, J. Biol. Chem. 267:16553-16560; Holvoet et al.,1995, Biochemistry 34:13334-13342; Sorci-Thomas et al., 1997, J. Biol.Chem. 272(11):7278-7284; and Frank et al., 1997, Biochemistry36:1798-1806).

In one study, Fukushima et al. synthesized a 22-residue peptide composedentirely of Glu, Lys and Leu residues arranged periodically so as toform an amphipathic α-helix with equal hydrophilic and hydrophobic faces("ELK peptide") (Fukushima et al., 1979, J. Amer. Chem. Soc.101(13):3703-3704; Fukushima et al., 1980, J. Biol. Chem.255:10651-10657). The ELK peptide shares 41% sequence homology with the198-219 fragment of ApoA-I. As studied by quantitative ultrafiltration,gel permeation chromatography and circular dichroism, this ELK peptidewas shown to effectively associate with phospholipids and mimic some ofthe physical and chemical properties of ApoA-I (Kaiser et al., 1983,Proc. Natl. Acad. Sci. U.S.A. 80:1137-1140; Kaiser et al., 1984, Science223:249-255; Fukushima et al., 1980, supra; Nakagawa et al., 1985, J.Am. Chem. Soc. 107:7087-7092). Yokoyama et al. concluded from suchstudies that the crucial factor for LCAT activation is simply thepresence of a large enough amphipathic structure (Yokoyama et al., 1980,J. Biol. Chem. 255(15):7333-7339). A dimer of this 22-residue peptidewas later found to more closely mimic ApoA-I than the monomer; based onthese results, it was suggested that the 44-mer, which is punctuated inthe middle by a helix breaker (either Gly or Pro), represented theminimal functional domain in ApoA-I (Nakagawa et al., 1985, supra).

Another study involved model amphipathic peptides called "LAP peptides"(Pownall et al., 1980, Proc. Natl. Acad. Sci. U.S.A. 77(6):3154-3158;Sparrow et al., 1981, In: Peptides: Synthesis-Structure-Function, Rochand Gross, Eds., Pierce Chem. Co., Rockford, Ill., 253-256). Based onlipid binding studies with fragments of native apolipoproteins, severalLAP peptides were designed, named LAP-16, LAP-20 and LAP-24 (containing16, 20 and 24 amino acid residues, respectively). These modelamphipathic peptides share no sequence homology with the apolipoproteinsand were designed to have hydrophilic faces organized in a manner unlikethe class A-type amphipathic helical domains associated withapolipoproteins (Segrest et al., 1992, J. Lipid Res. 33:141-166). Fromthese studies, the authors concluded that a minimal length of 20residues is necessary to confer lipid-binding properties to modelamphipathic peptides.

Studies with mutants of LAP20 containing a proline residue at differentpositions in the sequence indicated that a direct relationship existsbetween lipid binding and LCAT activation, but that the helicalpotential of a peptide alone does not lead to LCAT activation (Ponsin etal., 1986 J. Biol. Chem. 261(20):9202-9205). Moreover, the presence ofthis helix breaker (Pro) close to the middle of the peptide reduced itsaffinity for phospholipid surfaces as well as its ability to activateLCAT. While certain of the LAP peptides were shown to bind phospholipids(Sparrow et al., supra), controversy exists as to the extent to whichLAP peptides are helical in the presence of lipids (Buchko et al., 1996,J. Biol. Chem. 271(6):3039-3045; Zhong et al., 1994, Peptide Research7(2):99-106).

Segrest et al. have synthesized peptides composed of 18 to 24 amino acidresidues that share no sequence homology with the helices of ApoA-I(Kannelis et al., 1980, J. Biol. Chem. 255(3):11464-11472; Segrest etal., 1983, J. Biol. Chem. 258:2290-2295). The sequences werespecifically designed to mimic the amphipathic helical domains of classA exchangeable apolipoproteins in terms of hydrophobic moment (Eisenberget al., 1982, Nature 299:371-374) and charge distribution (Segrest etal., 1990, Proteins 8:103-117; U.S. Pat. No. 4,643,988). One 18-residuepeptide, the "18A" peptide, was designed to be a model class-A α-helix(Segrest et al., 1990, supra). Studies with these peptides and otherpeptides having a reversed charged distribution, like the "18R" peptide,have consistently shown that charge distribution is critical foractivity; peptides with a reversed charge distribution exhibit decreasedlipid affinity relative to the 18A class-A mimics and a lower helicalcontent in the presence of lipids (Kanellis et al., 1980, J. Biol. Chem.255:11464-11472; Anantharamaiah et al., 1985, J. Biol. Chem.260:10248-10255; Chung et al., 1985, J. Biol. Chem. 260:10256-10262;Epand et al., 1987, J. Biol. Chem. 262:9389-9396; Anantharamaiah et al.,1991, Adv. Exp. Med. Biol. 285:131-140).

Other synthetic peptides sharing no sequence homology with theapolipoproteins which have been proposed with limited success includedimers and trimers of the 18A peptide (Anantharamaiah et al., 1986,Proteins of Biological Fluids 34:63-66), GALA and EALA peptides(Subbarao et al., 1988, PROTEINS: Structure, Function and Genetics3:187-198) and ID peptides (Labeur et al., 1997, Arteriosclerosis,Thrombosis and Vascular Biology 17:580-588) and the 18AM4 peptide(Brasseur et al., 1993, Biochim. Biophys. Acta 1170:1-7).

A "consensus" peptide containing 22-amino acid residues based on thesequences of the helices of human ApoA-I has also been designed(Anantharamaiah et al., 1990, Arteriosclerosis 10(1):95-105;Venkatachalapathi et al., 1991, Mol. Conformation and Biol.Interactions, Indian Acad. Sci. B:585-596). The sequence was constructedby identifying the most prevalent residue at each position of thehypothesized helices of human ApoA-I. Like the peptides described above,the helix formed by this peptide has positively charged amino acidresidues clustered at the hydrophilic-hydrophobic interface, negativelycharged amino acid residues clustered at the center of the hydrophilicface and a hydrophobic angle of less than 180°. While a dimer of thispeptide is somewhat effective in activating LCAT, the monomer exhibitedpoor lipid binding properties (Venkatachalapathi et al., 1991, supra).

Based primarily on in vitro studies with the peptides described above, aset of "rules" has emerged for designing peptides which mimic thefunction of apoA-I. Significantly, it is thought that an amphipathicα-helix having positively charged residues clustered at thehydrophilic-hydrophobic interface and negatively charged amino acidresidues clustered at the center of the hydrophilic face is required forlipid affinity and LCAT activation (Venkatachalapathi et al., 1991,supra). Anantharamaiah et al. have also indicated that the negativelycharged Glu residue at position 13 of the consensus 22-mer peptide,which is positioned within the hydrophobic face of the α-helix, plays animportant role in LCAT activation (Anantharamaiah et al., 1991, supra).Furthermore, Brasseur has indicated that a hydrophobic angle (pho angle)of less than 180° is required for optimal lipid-apolipoprotein complexstability, and also accounts for the formation of discoidal particleshaving the peptides around the edge of the lipid bilayer (Brasseur,1991, J. Biol. Chem. 66(24):16120-16127). Rosseneu et al. have alsoinsisted that a hydrophobic angle of less than 180° is required for LCATactivation (WO93/25581).

However, despite these "rules" to date, no one has designed or produceda peptide as active as ApoA-I--the best having less than 40% of theactivity of ApoA-I as measured by the LCAT activation assay describedherein. None of the peptide "mimetics" described in the literature havebeen demonstrated to be useful as a drug.

In view of the foregoing, there is a need for the development of astable ApoA-I agonist that mimics the activity of ApoA-I and which isrelatively simple and cost-effective to produce. However, the "rules"for designing efficacious ApoA-I mimetics have not been unraveled andthe principles for designing organic molecules with the function ofApoA-I are unknown.

3. SUMMARY OF THE INVENTION

The invention relates to ApoA-I agonists capable of forming amphipathicα-helices that mimic the activity of ApoA-I, with specific activities,i.e., units of activity (activation of LCAT)/unit of mass), approachingor exceeding that of the native molecule. In particular, the ApoA-Iagonists of the invention are peptides or peptide analogues that: formamphipathic helices (in the presence of lipids), bind lipids, formpre-β-like or HDL-like complexes, activate LCAT, increase serum levelsof HDL fractions, and promote cholesterol efflux.

The invention is based, in part, on the applicants' design and discoveryof peptides that mimic the function of ApoA-I. The peptides of theinvention were designed based on the supposed helical structure andamphipathic properties of the 22 amino acid consensus sequence which wasderived from the helical repeats of ApoA-I. Surprisingly, the peptidesof the invention have a specific activity well above that reported forApoA-I-derived peptides described in the literature. Indeed, someembodiments of the invention approach 100% of the activity of nativeApoA-I, whereas superagonists described herein exceed the specificactivity of ApoA-I.

One aspect of the invention is also based, in part, on the Applicants'discovery that amino acid residues 14-17 of Segrest's consensus 22-merscould be deleted without loss of LCAT activation. In some cases, thedeletion of amino acid residues enhances LCAT activation. In anotheraspect of the invention, other residues of the consensus sequence canalso be deleted with enhanced activity. In addition, in someembodiments, the deletion can be used in conjunction with alteration ofamino acid residues. In some preferred embodiments, deletion of 4residues from Segrest's consensus 22-mer peptide is used in conjunctionwith changes at residue 5, 9 and 13 to a hydrophobic leucine.

The invention is illustrated by way of working examples that describethe structure, preparation and use of particular amphipathic peptidesthat form helices (in the presence of lipids), bind lipids, formcomplexes and increase LCAT activity. Based upon the structure andactivity of the exemplified embodiments, the applicants have devised aset of "rules" which can be used to design altered or mutated forms thatare also within the scope of the invention.

The invention also relates to pharmaceutical formulations containingsuch ApoA-I agonists (either as peptides or peptide-lipid complexes) asthe active ingredient, as well as methods for preparing suchformulations and their use to treat diseases associated withdyslipoproteinemia (e.g., cardiovascular diseases, atherosclerosis,metabolic syndrome), restenosis, or endotoxemia (e.g., septic shock).

3.1. Abbreviations

As used herein, the abbreviations for the genetically encodedL-enantiomeric amino acids are conventional and are as follows:

    ______________________________________                                                        One-Letter                                                                             Common                                               Amino Acid      Symbol   Abbreviation                                         ______________________________________                                        Alanine         A        Ala                                                  Arginine        R        Arg                                                  Asparagine      N        Asn                                                  Aspartic acid   D        Asp                                                  Cysteine        C        Cys                                                  Glutamine       Q        Gln                                                  Glutamic acid   E        Glu                                                  Glycine         G        Gly                                                  Histidine       H        His                                                  Isoleucine      I        Ile                                                  Leucine         L        Leu                                                  Lysine          K        Lys                                                  Methionine      M        Met                                                  Phenylalanine   F        Phe                                                  Proline         P        Pro                                                  Serine          S        Ser                                                  Threonine       T        Thr                                                  Tryptophan      W        Trp                                                  Tyrosine        Y        Tyr                                                  Valine          V        Val                                                  ______________________________________                                    

The abbreviations used for the D-enantiomers of the genetically encodedamino acids are lower-case equivalents of the one-letter symbols. Forexample, "R" designates L-arginine and "r" designates D-arginine.

3.2. Definitions

As used herein, the following terms shall have the following meanings:

"Alkyl:" refers to a saturated branched, straight chain or cyclichydrocarbon radical. Typical alkyl groups include, but are not limitedto, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl,isopentyl, hexyl, and the like. In preferred embodiments, the alkylgroups are (C₁ -C₆) alkyl.

"Alkenyl:" refers to an unsaturated branched, straight chain or cyclichydrocarbon radical having at least one carbon-carbon double bond. Theradical may be in either the cis or trans conformation about the doublebond(s). Typical alkenyl groups include, but are not limited to,ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, tert-butenyl,pentenyl, hexenyl and the like. In preferred embodiments, the alkenylgroup is (C₁ -C₆) alkenyl.

"Alkynyl:" refers to an unsaturated branched, straight chain or cyclichydrocarbon radical having at least one carbon-carbon triple bond.Typical alkynyl groups include, but are not limited to, ethynyl,propynyl, butynyl, isobutynyl, pentynyl, hexynyl and the like. Inpreferred embodiments, the alkynyl group is (C₁ -C₆) alkynyl.

"Aryl:" refers to an unsaturated cyclic hydrocarbon radical having aconjugated π electron system. Typical aryl groups include, but are notlimited to, penta-2,4-diene, phenyl, naphthyl, anthracyl, azulenyl,chrysenyl, coronenyl, fluoranthenyl, indacenyl, idenyl, ovalenyl,perylenyl, phenalenyl, phenanthrenyl, picenyl, pleiadenyl, pyrenyl,pyranthrenyl, rubicenyl, and the like. In preferred embodiments, thearyl group is (C₅ -C₂₀) aryl, with (C₅ -C₁₀) being particularlypreferred.

"Alkaryl:" refers to a straight-chain alkyl, alkenyl or alkynyl groupwherein one of the hydrogen atoms bonded to a terminal carbon isreplaced with an aryl moiety. Typical alkaryl groups include, but arenot limited to, benzyl, benzylidene, benzylidyne, benzenobenzyl,naphthenobenzyl and the like. In preferred embodiments, the alkarylgroup is (C₆ -C₂₆) alkaryl, i.e., the alkyl, alkenyl or alkynyl moietyof the alkaryl group is (C₁ -C₆) and the aryl moiety is (C₅ -C₂₀). Inparticularly preferred embodiments, the alkaryl group is (C₆ -C₁₃)alkaryl, i.e., the alkyl, alkenyl or alkynyl moiety of the alkaryl groupis (C₁ -C₃) and the aryl moiety is (C₅ -C₁₀).

"Heteroaryl:" refers to an aryl moiety wherein one or more carbon atomsis replaced with another atom, such as N, P, O, S, As, Se, Si, Te, etc.Typical heteroaryl groups include, but are not limited to, acridarsine,acridine, arsanthridine, arsindole, arsindoline, carbazole, β-carboline,chromene, cinnoline, furan, imidazole, indazole, indole, indolizine,isoarsindole, isoarsinoline, isobenzofuran, isochromene, isoindole,isophosphoindole, isophosphinoline, isoquinoline, isothiazole,isoxazole, naphthyridine, perimidine, phenanthridine, phenanthroline,phenazine, phosphoindole, phosphinoline, phthalazine, pteridine, purine,pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole,pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,selenophene, tellurophene, thiophene and xanthene. In preferredembodiments, the heteroaryl group is a 5-20 membered heteroaryl, with5-10 membered aryl being particularly preferred.

"Alkheteroaryl:" refers to a straight-chain alkyl, alkenyl or alkynylgroup where one of the hydrogen atoms bonded to a terminal carbon atomis replaced with a heteroaryl moiety. In preferred embodiments, thealkheteroaryl group is 6-26 membered alkheteroaryl, i.e., the alkyl,alkenyl or alkynyl moiety of the alkheteroaryl is (C₁ -C₆) and theheteroaryl is a 5-20-membered heteroaryl. In particularly preferredembodiments the alkheteroaryl is 6-13 membered alkheteroaryl, i.e., thealkyl, alkenyl or alkynyl moiety is a 5-10 membered heteroaryl.

"Substituted Alkyl, Alkenyl, Alkynyl, Aryl, Alkaryl, Heteroaryl orAlkheteroaryl:" refers to an alkyl, alkenyl, alkynyl, aryl, alkaryl,heteroaryl or alkheteroaryl group in which one or more hydrogen atoms isreplaced with another substituent. Preferred substituents include --OR,--SR, --NRR, --NO₂, --CN, halogen, --C(O)R, --C(O)OR and --C(O)NR, whereeach R is independently hydrogen, alkyl, alkenyl, alkynyl, aryl,alkaryl, heteroaryl or alkheteroaryl.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A is a Schiffer-Edmundson helical wheel diagram of an idealizedamphipathic α-helix in which open circles represent hydrophilic aminoacid residues and shaded circles represent hydrophobic amino acidresidues.

FIG. 1B is a helical net diagram of the idealized amphipathic helix ofFIG. 1A.

FIG. 1C is a helical cylinder diagram of the idealized amphipathic helixof FIG. 1A.

FIG. 2A is a Schiffer-Edmundson helical wheel diagram of the corepeptide of structure (I) illustrating the amphipathicity of the helix(open circles represent hydrophilic amino acid residues, shaded circlesrepresent hydrophobic amino acid residues and partially shaded circlesrepresent either hydrophilic or hydrophobic amino acid residues).

FIG. 2B is a helical net diagram of the core peptide of structure (I)illustrating the hydrophobic face of the helix.

FIG. 2C is a helical net diagram of the core peptide of structure (I)illustrating the hydrophilic face of the helix.

FIG. 3A is a helical net diagram illustrating the hydrophilic face ofSegrest's 18A peptide (DWLKAFYDKVAEKLKEAF; SEQ ID NO:244).

FIG. 3B is a helical net diagram illustrating the hydrophilic face ofexemplary core peptide 210 (PVLDLFRELLEELKQKLK; SEQ ID NO:210).

FIG. 3C is a helical net diagram illustrating the hydrophilic face ofSegrest's consensus 22-mer peptide (PVLDEFREKLNEELEALKQKLK; SEQ IDNO:75).

FIG. 4A is a helical net diagram illustrating the hydrophobic face ofSegrest's 18A peptide (SEQ ID NO:244).

FIG. 4B is a helical net diagram illustrating the hydrophobic face ofexemplary core peptide 210 (SEQ ID NO:210).

FIG. 4C is a helical net diagram illustrating the hydrophobic face ofSegrest's consensus 22-mer peptide (SEQ ID NO:75).

FIG. 5A is a Schiffer-Edmundson helical wheel diagram of Segrest's 18Apeptide (SEQ ID NO:244).

FIG. 5B is a Schiffer-Edmundson helical wheel diagram of exemplary corepeptide 210 (SEQ ID NO:210).

FIG. 6A illustrates a tertiary-order branched network of the invention.

FIG. 6B illustrates a quaternary-order branched network of theinvention.

FIG. 6C illustrates a mixed-order branched network of the invention.

FIG. 6D illustrates exemplary "Lys-tree" branched networks of theinvention.

FIG. 7A is a graph illustrating the differences between the observed Hαchemical shifts and the tabulated random coil Hα chemical shifts forpeptide 210 (SEQ ID NO:210) and Segrest's consensus 22-mer peptide (SEQID NO:75).

FIG. 7B is a graph illustrating the differences between the observedamide proton chemical shifts and the tabulated random coil amide protonchemical shifts for peptide 210 (SEQ ID NO:210) and Segrest's consensus22-mer peptide (SEQ ID NO:75).

FIG. 8A is a cartoon depicting the various aggregation states andpeptide-lipid complexes that can be obtained with the ApoA-I agonists ofthe invention. Left: Multimerization process of the peptides resultingfrom the interaction of several peptide helices and leading to theformation of oligomers in conditions of defined peptide concentration,pH and ionic strength. Center: The interaction of the peptides (in anyof these states of aggregation) with lipidic entities (such as SUVs)leads to lipid reorganization. Right: By changing the lipid:peptidemolar ratio, different types of peptide-lipid complexes can be obtained,from lipid-peptide comicelles at low lipid-peptide ratios, to discoidalparticles and finally to large multilamellar complexes at increasinglyhigher lipid:peptide ratios.

FIG. 8B illustrates the generally-accepted model for discoidalpeptide-lipid complexes formed in a defined range of lipid:peptideratios. Each peptide surrounding the disc edge is in close contact withits two nearest neighbors.

5. DETAILED DESCRIPTION OF THE INVENTION

The ApoA-I agonists of the invention mimic ApoA-I function and activity.They form amphipathic helices (in the presence of lipids), bind lipids,form pre-β-like or HDL-like complexes, activate LCAT, increase serum HDLconcentration and promote cholesterol efflux. The biological function ofthe peptides correlates with their helical structure, or conversion tohelical structures in the presence of lipids.

The ApoA-I agonists of the invention can be prepared in stable bulk orunit dosage forms, e.g., lyophilized products, that can be reconstitutedbefore use in vivo or reformulated. The invention includes thepharmaceutical formulations and the use of such preparations in thetreatment of hyperlipidemia, hypercholesterolemia, coronary heartdisease, atherosclerosis, and other conditions such as endotoxemiacausing septic shock.

The invention is illustrated by working examples which demonstrate thatthe ApoA-I agonists of the invention are extremely efficient atactivating LCAT, and thus promote RCT. Use of the ApoA-I agonists of theinvention in vivo in animal models results in an increase in serum HDLconcentration.

The invention is set forth in more detail in the subsections below,which describe: the composition and structure of the ApoA-I peptideagonists; structural and functional characterization; methods ofpreparation of bulk and unit dosage formulations; and methods of use.

5.1. Peptide Structure and Function

The ApoA-I agonists of the invention are generally peptides, oranalogues thereof, which are capable of forming amphipathic α-helices inthe presence of lipids and which mimic the activity of ApoA-I. Theagonists have as their main feature a "core" peptide composed of 15 to22 amino acid residues, preferably 18 amino acid residues, or ananalogue thereof wherein at least one amide linkage in the peptide isreplaced with a substituted amide, an isostere of an amide or an amidemimetic.

The ApoA-I agonists of the invention are based, in part, on theapplicants' surprising discovery that altering and/or deleting certainamino acid residues in the primary sequence of the 22-mer consensussequence of Venkatachalapathi et al., 1991, Mol. Conformation and Biol.Interactions, Indian Acad. Sci. B:585-596 (PVLDEFREKLNEELEALKQKLK; SEQID NO:75; hereinafter "Segrest's consensus 22-mer" or "consensus22-mer") that were thought to be critical for activity yields syntheticpeptides which exhibit activities that approach, or in some embodimentseven exceed, the activity of native ApoA-I. In one aspect of theinvention, four amino acid residues of the consensus 22-mer, such asresidues 14-17, are deleted to form 18-mers capable of LCAT activation.In other additional embodiments, four other residues are deleted. Insome embodiments, three charged amino acid residues that were thought tobe critical for activity (Glu-5, Lys-9 and Glu-13) are replaced with ahydrophobic residue such as Leu.

While not intending to be bound by any particular theory, it is believedthat the helix formed by the ApoA-I agonists of the invention moreclosely mimics the structural and functional properties of theamphipathic helical regions of native ApoA-I that are important foreffecting lipid-binding, cholesterol efflux and LCAT activation thandoes the α-helix formed by the ApoA-I mimetic peptides described in theliterature, thereby resulting in peptides that exhibit significantlyhigher ApoA-I-like activity than these other peptides. Indeed, whereasmany of the ApoA-I agonists of the invention approach, and in someembodiments even exceed, the activity of ApoA-I, to date, the bestpeptide ApoA-I mimics described in the literature--peptide 18AM4(EWLEAFYKKVLEKLKELF; SEQ ID NO:246) (Corinjn et al., 1993, Biochim.Biophys. Acta 1170:8-16; Labeur et al., October 1994, Arteriosclerosis:Abstract Nos. 186 and 187) and N-acetylated, C-amidated peptide 18AM4(SEQ ID NO: 239) (Brasseur, 1993, Biochim. Biophys. Acta1170:1-7)--exhibit less than 4% and 11%, respectively, of the activityof ApoA-I as measured by the LCAT activation assay described herein.

In one illustrative embodiment of the invention, the core peptides (oranalogues thereof) that compose the ApoA-I agonists of the inventionhave the following structural formula (I):

    X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18

wherein:

X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);

X₂ is an aliphatic amino acid;

X₃ is Leu (L);

X₄ is an acidic amino acid;

X₅ is Leu (L) or Phe (F);

X₆ is Leu (L) or Phe (F);

X₇ is a basic amino acid;

X₈ is an acidic amino acid;

X₉ is Leu (L) or Trp (W);

X₁₀ is Leu (L) or Trp (W);

X₁₁ is an acidic amino acid or Asn (N);

X₁₂ is an acidic amino acid;

X₁₃ is Leu (L), Trp (W) or Phe (F);

X₁₄ is a basic amino acid or Leu (L);

X₁₅ is Gln (Q) or Asn (N);

X₁₆ is a basic amino acid;

X₁₇ is Leu (L); and

X₁₈ is a basic amino acid.

The core peptides of structure (I) are defined, in part, in terms ofamino acids of designated classes. The definitions of the variousdesignated classes are provided infra in connection with the descriptionof mutated or altered embodiments of structure (I).

In the core peptides of structure (I), the symbol "-" between amino acidresidues X_(n) generally designates a backbone constitutive linkingfunction. Thus, the symbol "-" usually represents a peptide bond oramide linkage (--C(O)NH--). It is to be understood, however, that thepresent invention contemplates peptide analogues wherein one or moreamide linkages is optionally replaced with a linkage other than amide,preferably a substituted amide or an isostere of amide. Thus, while thevarious X_(n) residues within structure (I) are generally described interms of amino acids, and preferred embodiments of the invention areexemplified by way of peptides, one having skill in the art willrecognize that in embodiments having non-amide linkages, the term "aminoacid" or "residue" as used herein refers to other bifunctional moietiesbearing groups similar in structure to the side chains of the aminoacids.

Substituted amides generally include, but are not limited to, groups ofthe formula --C(O)NR--, where R is (C₁ -C₆) alkyl, substituted (C₁ -C₆)alkyl, (C₁ -C₆) alkenyl, substituted (C₁ -C₆) alkenyl, (C₁ -C₆) alkynyl,substituted (C₁ -C₆) alkynyl, (C₅ -C₂₀) aryl, substituted (C₅ -C₂₀)aryl, (C₆ -C₂₆) alkaryl, substituted (C₆ -C₂₆) alkaryl, 5-20 memberedheteroaryl, substituted 5-20 membered heteroaryl or 6-26 memberedalkheteroaryl and substituted 6-26 membered alkheteroaryl.

Isosteres of amide generally include, but are not limited to, --CH₂NH--, --CH₂ S--, --CH₂ CH₂ --, --CH═CH-- (cis and trans), --C(O)CH₂ --,--CH(OH)CH₂ -- and --CH₂ SO--. Compounds having such non-amide linkagesand methods for preparing such compounds are well-known in the art (see,e.g., Spatola, March 1983, Vega Data Vol. 1, Issue 3; Spatola, 1983,"Peptide Backbone Modifications" In: Chemistry and Biochemistry of AminoAcids Peptides and Proteins, Weinstein, ed., Marcel Dekker, New York, p.267 (general review); Morley, 1980, Trends Pharm. Sci. 1:463-468; Hudsonet al., 1979, Int. J. Prot. Res. 14:177-185 (--CH₂ NH--, --CH₂ CH₂ --);Spatola et al., 1986, Life Sci. 38:1243-1249 (--CH₂ --S); Hann, 1982, J.Chem. Soc. Perkin Trans. I. 1:307-314 (--CH═CH--, cis and trans);Almquist et al., 1980, J. Med. Chem. 23:1392-1398 (--COCH₂ --);Jennings-White et al., Tetrahedron. Lett. 23:2533 (--COCH₂ --); EuropeanPatent Application EP 45665 (1982) CA 97:39405 (--CH(OH)CH₂ --);Holladay et al., 1983, Tetrahedron Lett. 24:4401-4404 (--C(OH)CH₂ --);and Hruby, 1982, Life Sci. 31:189-199 (--CH₂ --S--)--.

Additionally, one or more amide linkages can be replaced withpeptidomimetic or amide mimetic moieties which do not significantlyinterfere with the structure or activity of the peptides. Suitable amidemimetic moieties are described, for example, in Olson et al., 1993, J.Med. Chem. 36:3039-3049.

A critical feature of the core peptides of structure (I), is theirability to form an amphipathic α-helix in the presence of lipids. Byamphipathic is meant that the α-helix has opposing hydrophilic andhydrophobic faces oriented along its long axis, i.e., one face of thehelix projects mainly hydrophilic side chains while the opposite faceprojects mainly hydrophobic side chains. FIGS. 1A and 1B present twoillustrative views of the opposing hydrophilic and hydrophobic faces ofan exemplary idealized amphipathic α-helix. FIG. 1A is aSchiffer-Edmundson helical wheel diagram (Schiffer and Edmundson, 1967,Biophys. J. 7:121-135). In the wheel, the long axis of the helix isperpendicular to the page. Starting with the N-terminus, successiveamino acid residues (represented by circles) are radially distributedabout the perimeter of a circle at 100° intervals. Thus, amino acidresidue n+1 is positioned 100° from residue n, residue n+2 is positioned100° from residue n+1, and so forth. The 100° placement accounts for the3.6 amino acid residues per turn that are typically observed in anidealized α-helix. In FIG. 1A, the opposing hydrophilic and hydrophobicfaces of the helix are clearly visible; hydrophilic amino acids arerepresented as open circles and hydrophobic amino acid residues arerepresented as shaded circles.

FIG. 1B presents a helical net diagram of the idealized amphipathichelix of FIG. 1A. (Lim, 1978, FEBS Lett. 89:10-14). In a typical helicalnet diagram, the α-helix is presented as a cylinder that has been cutalong the center of its hydrophilic face and flattened. Thus, the centerof the hydrophobic face, determined by the hydrophobic moment of thehelix (Eisenberg et al., 1982, Nature 299:371-374), lies in the centerof the figure and is oriented so as to rise out of the plane of thepage. An illustration of the helical cylinder prior to being cut andflattened is depicted in FIG. 1C. By cutting the cylinder alongdifferent planes, different views of the same amphipathic helix can beobserved, and different information about the properties of the helixobtained.

The amphipathic nature of the α-helix formed by the core peptides ofstructure (I) in the presence of lipids is illustrated in FIG. 2. FIG.2A presents a Schiffer-Edmundson helical wheel diagram, FIG. 2B presentsa helical net diagram illustrating the hydrophobic face and FIG. 2Cpresents a helical net diagram illustrating the hydrophilic face. Ineach of FIGS. 2A, 2B and 2C, hydrophilic residues are represented asopen circles, hydrophobic residues as shaded circles, and residues whichcan be either hydrophilic or hydrophobic as partially shaded circles. Aswill be discussed more thoroughly below in conjunction with altered ormutated forms of the peptides of structure (I), certain amino acidresidues can be replaced with other amino acid residues such that thehydrophilic and hydrophobic faces of the helix formed by the peptidesmay not be composed entirely of hydrophilic and hydrophobic amino acids,respectively. Thus, it is to be understood that when referring to theamphipathic α-helix formed by the core peptides of the invention, thephrase "hydrophilic face" refers to a face of the helix having overallnet hydrophilic character. The phrase "hydrophobic face" refers to aface of the peptide having overall net hydrophobic character.

While not intending to be bound by any particular theory, it is believedthat certain structural and/or physical properties of the amphipathichelix formed by the core peptides of structure (I), are important foractivity. These properties include the degree of amphipathicity, overallhydrophobicity, mean hydrophobicity, hydrophobic and hydrophilic angles,hydrophobic moment, mean hydrophobic moment, and net charge of theα-helix.

While the helical wheel diagrams of FIG. 2A provide a convenient meansof visualizing the amphipathic nature of the core peptides of structure(I), the degree of amphipathicity (degree of asymmetry ofhydrophobicity) can be conveniently quantified by calculating thehydrophobic moment (μ_(H)) of the helix. Methods for calculating μ_(H)for a particular peptide sequence are well-known in the art, and aredescribed, for example in Eisenberg, 1984, Ann. Rev. Biochem.53:595-623. The actual μ_(H) obtained for a particular peptide willdepend on the total number of amino acid residues composing the peptide.Thus, it is generally not informative to directly compare μ_(H) forpeptides of different lengths.

The amphipathicities of peptides of different lengths can be directlycompared by way of the mean hydrophobic moment (<μ_(H) >). The meanhydrophobic moment can be obtained by dividing μ_(H) by the number ofresidues in the helix (i.e., <μ_(H) >=μ_(H) /N). Generally, corepeptides which exhibit a <μ_(H) > in the range of 0.55 to 0.65, asdetermined using the normalized consensus hydrophobicity scale ofEisenberg (Eisenberg, 1984, J. Mol. Biol. 179:125-142) are considered tobe within the scope of the present invention, with a <μ_(H) > in therange of 0.58 to 0.62 being preferred.

The overall or total hydrophobicity (H_(o)) of a peptide can beconveniently calculated by taking the algebraic sum of thehydrophobicities of each amino acid residue in the peptide ##EQU1##where N is the number of amino acid residues in the peptide and H_(i) isthe hydrophobicity of the ith amino acid residue). The meanhydrophobicity (<H_(o) >) is the hydrophobicity divided by the number ofamino acid residues (i.e., <H_(o) >=H_(o) /N). Generally, core peptidesthat exhibit a mean hydrophobicity in the range of -0.150 to -0.070, asdetermined using the normalized consensus hydrophobicity scale ofEisenberg (Eisenberg, 1984, J. Mol. Biol. 179:125-142) are considered tobe within the scope of the present invention, with a mean hydrophobicityin the range of -0.130 to -0.050 being preferred.

The total hydrophobicity of the hydrophobic face (H_(o) ^(pho)) of anamphipathic helix can be obtained by taking the sum of thehydrophobicities of the hydrophobic amino acid residues which fall intothe hydrophobic angle as defined below ##EQU2## where H_(i) is aspreviously defined and N_(H) is the total number of hydrophobic aminoacids in the hydrophobic face). The mean hydrophobicity of thehydrophobic face (<H_(o) ^(pho) >) is H_(o) ^(pho) /N_(H) where N_(H) isas defined above. Generally, core peptides which exhibit a <H_(o)^(pho) > in the range of 0.90 to 1.20, as determined using the consensushydrophobicity scale of Eisenberg (Eisenberg, 1984, supra; Eisenberg etal., 1982, supra) are considered to be within the scope of the presentinvention, with a <H_(o) ^(pho) > in the range of 0.950 to 1.10 beingpreferred.

The hydrophobic angle (pho angle) is generally defined as the angle orarc covered by the longest continuous stretch of hydrophobic amino acidresidues when the peptide is arranged in the Schiffer-Edmundson helicalwheel representation (i.e., the number of contiguous hydrophobicresidues on the wheel multiplied by 20°). The hydrophilic angle (phiangle) is the difference between 360° and the pho angle (i.e., 360°-phoangle). Those of skill in the art will recognize that the pho and phiangles will depend, in part, on the number of amino acid residues in thepeptide. For example, referring to FIGS. 5A and 5B, it can be seen thatonly 18 amino acids fit around one rotation of the Schiffer-Edmundsonhelical wheel. Fewer amino acids leave a gap in the wheel; more aminoacids cause certain positions of the wheel to be occupied by more thanone amino acid residue.

In the case of peptides containing more than 18 amino acid residues, by"continuous" stretch of hydrophobic amino acid residues is meant that atleast one amino acid residue at positions along the wheel occupied bytwo or more amino acids is a hydrophobic amino acid.

Typically, core peptides composed of 18 or fewer amino acids having apho angle in the range of 120° to 160° are considered to be within thescope of the invention, with a pho angle in the range of 130° to 150°being preferred. Embodiments containing more than 18 amino acidstypically have a pho angle in the range of 160° to 220°, with 180° to200° being preferred.

Certain structural and/or physical characteristics of the core peptidesof structure (I) are illustrated in FIGS. 3 and 4. FIG. 3B presents ahelical net diagram of an exemplary core peptide of the invention,peptide 210 (PVLDLFRELLEELKQKLK; SEQ ID NO:210), illustrating the chargedistribution along the hydrophilic face of the helix. In FIG. 3B, thehelical cylinder has been cut along the center of the hydrophobic faceand flattened. The three hydrophobic Leu (L) residues that replacehydrophilic residues in Segrest's consensus 22-mer (residues 5, 9 and13) (FIG. 3C) are shaded. As can be seen in FIG. 3B, positively-chargedamino acid residues are clustered at the last C-terminal turn of thehelix (the C-terminus is at the top of the page). While not intending tobe bound by any particular theory, it is believed that this cluster ofbasic residues at the C-terminus (residues 14, 16 and 18) stabilizes thehelix through charge (NH₃ ⁺)-helix dipole electrostatic interactions. Itis also thought that stabilization occurs through hydrophobicinteractions between lysine side chains and the helix core (see, Groebkeet al., 1996, Proc. Natl. Acad. Sci. U.S.A. 93:4025-4029; Esposito etal., 1997, Biopolymers 41:27-35).

With the exception of the positively-charged C-terminal cluster(residues 14, 16 and 18), negative charges are distributed on the restof the hydrophilic face, with at least one negatively charged (acidic)amino acid residue per turn, resulting in a continuous stretch ofnegative charges along the hydrophilic face of the helix.

FIG. 4B presents a helical net diagram illustrating the hydrophobic faceof the amphipathic helix formed by exemplary core peptide 210 (SEQ IDNO:210). In FIG. 4B, the helical cylinder is cut along the center of thehydrophilic face and flattened. The hydrophobic face of the core peptideconsists of two hydrophobic residues per turn, except for the lastC-terminal turn, where basic residues dominate. NMR studies indicatethat amino acid residues 3, 6, 9 and 10 of an analogue 22-mer peptide(PVLDLFRELLNELLEALKQKLK; SEQ ID NO:4) form a hydrophobic cluster nearthe N-terminus of the helix. Phe-6 is centered in this cluster and isbelieved to play an important role in stabilizing the hydrophobiccluster.

While not intending to be bound by any particular theory, it is believedthat the hydrophobic cluster formed by residues 3, 6, 9 and 10 issignificant in effecting lipid binding and LCAT activation. Amphipathicpeptides are expected to bind phospholipids by pointing theirhydrophobic faces towards the alkyl chains of the lipid moieties. Thus,it is believed that this highly hydrophobic cluster contributes to thestrong lipid affinities observed for the core peptides of the invention.Since lipid binding is a prerequisite for LCAT activation, it isbelieved that this hydrophobic cluster is also essential for LCATactivation.

Aromatic residues are often found to be important in anchoring peptidesand proteins to lipids (De Kruijff, 1990, Biosci. Rep. 10:127-130;O'Neil and De Grado, 1990, Science 250:645-651; Blondelle et al., 1993,Biochim. Biophys. Acta 1202:331-336). Thus, it is further believed thatPhe-6, which is positioned at the center of the hydrophobic cluster, mayalso play a key role in anchoring the core peptides of structure (I) tolipids.

Interactions between the core peptides of the invention and lipids leadto the formation of peptide-lipid complexes. As illustrated in FIG. 8A,the type of complex obtained (comicelles, discs, vesicles ormultilayers) depends on the lipid:peptide molar ratio, with comicellesgenerally being formed at low lipid:peptide molar ratios and discoidaland vesicular or multilayer complexes being formed with increasinglipid:peptide molar ratios. This characteristic has been described foramphipathic peptides (Epand, The Amphipathic Helix, 1993) and for ApoA-I(Jones, 1992, Structure and Function of Apolipoproteins, Chapter 8, pp.217-250). The lipid:peptide molar ratio also determines the size andcomposition of the complexes (see, Section 5.3.1, infra).

The long axis of the α-helix formed by the core peptides of structure(I) has an overall curved shape. In typical amphipathic helices, it hasbeen found that the lengths of the hydrogen bonds of the hydrophilic andhydrophobic faces vary such that the hydrophobic side of the helix isconcave (Barlow and Thornton, 1988, J. Mol. Biol. 201:601-619; Zhou etal., 1992, J. Am. Chem. Soc. 33:11174-11183; Gesell et al., 1997, J.Biomol. NMR 9:127-135). While not intending to be bound by theory, it isbelieved that the overall curvature of the hydrophobic face of the helixmay be important in binding discoidal complexes--a curved helix permitsthe peptide to "fit" better around the edges of discoidal particles,thereby increasing the stability of the peptide-disc complex.

In the generally accepted structural model of ApoA-I, the amphipathicα-helices are packed around the edge of the discoidal HDL (see, FIG.8B). In this model, the helices are assumed to be aligned with theirhydrophobic faces pointing towards the lipid acyl chains (Brasseur etal., 1990, Biochim. Biophys. Acta 1043:245-252). The helices arearranged in an antiparallel fashion, and a cooperative effect betweenthe helices is thought to contribute to the stability of the discoidalHDL complex (Brasseur et al., supra). It has been proposed that onefactor that contributes to the stability of the HDL discoidal complex isthe existence of ionic interactions between acidic and basic residuesresulting in the formation of intermolecular salt bridges or hydrogenbonds between residues on adjacent anti-parallel helices. In this model,the peptides are considered not as a single entity, but as ininteraction with at least two other neighboring peptide molecules (FIG.8B).

It is also generally accepted that intramolecular hydrogen bond or saltbridge formation between acidic and basic residues, respectively, atpositions i and i+3 of the helix stabilize the helical structure(Marqusee et al., 1985, Proc. Natl. Acad. Sci. U.S.A. 84(24):8898-8902).One positive charge is located at residue 7 potentially contributing tohelix stability by forming a salt bridge with an acidic residue one turnaway.

Thus, additional key features of the core peptides of structure (I) aretheir ability to form intermolecular hydrogen-bonds with one anotherwhen aligned in an antiparallel fashion with their hydrophobic facespointing in the same direction, such as would be the case when thepeptides are bound to lipids and also their ability to formintermolecular hydrogen bonds or salt bridges near the N- and C-terminiof the helix. It is believed that the ability of the core peptides ofstructure (I) to closely pack and ionically interact to form intra-and/or inter-molecular salt bridges and/or hydrogen bonds when bound tolipids in an antiparallel fashion is an important feature of the corepeptides of the invention.

The ability of the core peptides to form favorable intermolecularpeptide-peptide interactions is also thought to be of relevance in theabsence of lipids. The core peptides of the invention self-associate,due in part to their high <μ_(H) >, <H_(o) > and hydrophobic angle (see,TABLE I, infra). The self-association phenomenon depends on theconditions of pH, peptide concentration and ionic strength, and canresult in several states of association, from monomeric to severalmultimeric forms (FIG. 10A). The hydrophobic core of peptide aggregatesfavors hydrophobic interactions with lipids. The ability of the peptidesto aggregate even at very low concentrations may favor their binding tolipids. It is thought that in the core of the peptide aggregatespeptide-peptide interactions also occur and may compete withlipid-peptide interactions.

In addition to the above-described properties, other parameters arethought to be important for activity as well, including the total numberof hydrophobic residues, the total number of charged residues, and thenet charge of the peptides.

A summary of the preferred physical and structural properties of thecore peptides of structure (I) is provided in TABLE I, below:

                  TABLE I                                                         ______________________________________                                        PHYSICAL PROPERTIES OF PREFERRED                                              ApoA-I AGONISTS OF STRUCTURE (I)                                              PROPERTY     RANGE        PREFERRED RANGE                                     ______________________________________                                        % hydrophobic amino                                                                        40-70        50-60                                               acids                                                                         <H.sub.o >   -0.150 to -0.070                                                                           -0.130 to -0.050                                    <H.sub.o.sup.pho >                                                                         0.90-1.2     0.95-1.10                                           <μ.sub.H >                                                                              0.55-0.65    0.58-0.62                                           pho angle    120°-160°                                                                    130°-150°                             # positively 3-5          4                                                   charged amino acids                                                           # negatively 3-5          4                                                   charged amino acids                                                           net charge   -1 to +1     0                                                   hydrophobic cluster                                                                       positions 3, 6, 9, 10 are hydrophobic amino acids                 acidic cluster                                                                            at least 1 acidic amino acid per turn except                                  for last 5 C-terminal amino acids                                 basic cluster                                                                             at least 2 basic amino acids in last 5 C-                                     terminal amino acids                                              ______________________________________                                    

The properties of the amphipathic α-helices formed by the core peptidesof the invention differ significantly from the properties of class Aamphipathic α-helices, particularly the class A α-helix of Segrest's 18Aand consensus 22-mer peptides. These differences are illustrated withexemplary core peptide 210 (SEQ ID NO:210) in FIGS. 3-5.

Referring to FIGS. 4A-4C, it can be seen that the hydrophobic face ofpeptide 210 has much greater hydrophobic character than the hydrophobicface of Segrest's 18A peptide or consensus 22-mer. In particular,residues 9 and 13 (shaded region of FIG. 4B) are hydrophobic Leu (L)residues in peptide 210 (SEQ ID NO:210) as compared to charged residuesin the 18A peptide (SEQ ID NO:244) and the consensus 22-mer (SEQ IDNO:75). The replacement of these two charged residues in Segrest's 18Aand consensus 22-mer peptides with hydrophobic Leu (L) residues leads tosignificant differences in the amphipathicity, hydrophobicity, pho angleand other properties of the helix.

A comparison of the physical and structural properties of peptide 210(SEQ ID NO:210) and Segrest's 18A peptide (SEQ ID NO:244) and consensus22-mer peptide (SEQ ID NO:75) is provided in TABLE II, below:

                  TABLE II                                                        ______________________________________                                        COMPARISON OF PROPERTIES OF EXEMPLARY                                         CORE PEPTIDE 210 (SEQ ID NO:210) WITH                                         SEGREST'S CONSENSUS 22-MER (SEQ ID NO:75)                                     AND 18A PEPTIDE (SEQ ID NO:244)                                                                    CONSENSUS                                                PROPERTY   18A       22-MER     PEPTIDE 210                                   ______________________________________                                        # amino acids                                                                            18        22         18                                            # hydrophilic                                                                            9         13         9                                             amino acids                                                                   # hydrophobic                                                                            9         9          9                                             amino acids                                                                   % hydrophobic                                                                            50        41         50                                            amino acids                                                                   <H.sub.o > -0.43     -0.293     -0.125                                        <H.sub.o.sup.pho >                                                                       0.778     0.960      1.081                                         <μ.sub.H >                                                                            0.485     0.425      0.597                                         pho angle  100°                                                                             100°                                                                              140°                                   # positively                                                                             4         5          4                                             charged amino                                                                 acids                                                                         # negatively                                                                             4         6          4                                             charged amino                                                                 acids                                                                         net charge 0         -1         0                                             ______________________________________                                    

These differences in properties lead to significant differences inactivity. Whereas Segrest's 18A peptide (SEQ ID NO:244) and consensus22-mer peptide (SEQ ID NO:75) exhibit only 5 and 10% LCAT activation,respectively, as compared with native ApoA-I in the assays describedherein, peptide 210 (SEQ ID NO:210) exhibits 46% activation as comparedwith native ApoA-I in the same assays. Peptide 193 (SEQ ID NO:193),which is the N-terminal acetylated and C-terminal amidated form ofpeptide 210, exhibits 96% LCAT activation in the same assay.

Certain amino acid residues in the core peptides of structure (I) can bereplaced with other amino acid residues without significantlydeleteriously affecting, and in many cases even enhancing, the activityof the peptides. Thus, also contemplated by the present invention arealtered or mutated forms of the core peptides of structure (I) whereinat least one defined amino acid residue in the structure is substitutedwith another amino acid residue. As one of the critical featuresaffecting the activity of the core peptides of the invention is believedto be their ability to form α-helices in the presence of lipids thatexhibit the amphipathic and other properties described above, it will berecognized that in preferred embodiments of the invention, the aminoacid substitutions are conservative, i.e., the replacing amino acidresidue has physical and chemical properties that are similar to theamino acid residue being replaced.

For purposes of determining conservative amino acid substitutions, theamino acids can be conveniently classified into two maincategories--hydrophilic and hydrophobic--depending primarily on thephysical-chemical characteristics of the amino acid side chain. Thesetwo main categories can be further classified into subcategories thatmore distinctly define the characteristics of the amino acid sidechains. For example, the class of hydrophilic amino acids can be furthersubdivided into acidic, basic and polar amino acids. The class ofhydrophobic amino acids can be further subdivided into apolar andaromatic amino acids. The definitions of the various categories of aminoacids that define structure (I) are as follows:

"Hydrophilic Amino Acid" refers to an amino acid exhibiting ahydrophobicity of less than zero according to the normalized consensushydrophobicity scale of Eisenberg et al., 1984, J. Mol. Biol.179:125-142. Genetically encoded hydrophilic amino acids include Thr(T), Ser (S), His (H), Glu (E), Asn (N), Gln (Q), Asp (D), Lys (K) andArg (R).

"Acidic Amino Acid" refers to a hydrophilic amino acid having a sidechain pK value of less than 7. Acidic amino acids typically havenegatively charged side chains at physiological pH due to loss of ahydrogen ion. Genetically encoded acidic amino acids include Glu (E) andAsp (D).

"Basic Amino Acid" refers to a hydrophilic amino acid having a sidechain pK value of greater than 7. Basic amino acids typically havepositively charged side chains at physiological pH due to associationwith hydronium ion. Genetically encoded basic amino acids include His(H), Arg (R) and Lys (K).

"Polar Amino Acid" refers to a hydrophilic amino acid having a sidechain that is uncharged at physiological pH, but which has at least onebond in which the pair of electrons shared in common by two atoms isheld more closely by one of the atoms. Genetically encoded polar aminoacids include Asn (N), Gln (Q) Ser (S) and Thr (T).

"Hydrophobic Amino Acid" refers to an amino acid exhibiting ahydrophobicity of greater than zero according to the normalizedconsensus hydrophobicity scale of Eisenberg, 1984, J. Mol. Biol.179:125-142. Genetically encoded hydrophobic amino acids include Pro(P), Ile (I), Phe (F), Val (V), Leu (L), Trp (W), Met (M), Ala (A), Gly(G) and Tyr (Y).

"Aromatic Amino Acid" refers to a hydrophobic amino acid with a sidechain having at least one aromatic or heteroaromatic ring. The aromaticor heteroaromatic ring may contain one or more substituents such as--OH, --SH, --CN, --F, --Cl, --Br, --I, --NO₂, --NO, --NH₂, --NHR,--NRR, --C(O)R, --C(O)OH, --C(O)OR, --C(O)NH₂, --C(O)NHR, --C(O)NRR andthe like where each R is independently (C₁ -C₆) alkyl, substituted (C₁-C₆) alkyl, (C₁ -C₆) alkenyl, substituted (C₁ -C₆) alkenyl, (C₁ -C₆)alkynyl, substituted (C₁ -C₆) alkynyl, (C₅ -C₂₀) aryl, substituted (C₅-C₂₀) aryl, (C₆ -C₂₆) alkaryl, substituted (C₆ -C₂₆) alkaryl, 5-20membered heteroaryl, substituted 5-20 membered heteroaryl, 6-26 memberedalkheteroaryl or substituted 6-26 membered alkheteroaryl. Geneticallyencoded aromatic amino acids include Phe (F), Tyr (Y) and Trp (W).

"Nonpolar Amino Acid" refers to a hydrophobic amino acid having a sidechain that is uncharged at physiological pH and which has bonds in whichthe pair of electrons shared in common by two atoms is generally heldequally by each of the two atoms (i.e., the side chain is not polar).Genetically encoded apolar amino acids include Leu (L), Val (V), Ile(I), Met (M), Gly (G) and Ala (A).

"Aliphatic Amino Acid" refers to a hydrophobic amino acid having analiphatic hydrocarbon side chain. Genetically encoded aliphatic aminoacids include Ala (A), Val (V), Leu (L) and Ile (I).

The amino acid residue Cys (C) is unusual in that it can form disulfidebridges with other Cys (C) residues or other sulfanyl-containing aminoacids. The ability of Cys (C) residues (and other amino acids with --SHcontaining side chains) to exist in a peptide in either the reduced free--SH or oxidized disulfide-bridged form affects whether Cys (C) residuescontribute net hydrophobic or hydrophilic character to a peptide. WhileCys (C) exhibits a hydrophobicity of 0.29 according to the normalizedconsensus scale of Eisenberg (Eisenberg, 1984, supra), it is to beunderstood that for purposes of the present invention Cys (C) iscategorized as a polar hydrophilic amino acid, notwithstanding thegeneral classifications defined above.

As will be appreciated by those of skill in the art, the above-definedcategories are not mutually exclusive. Thus, amino acids having sidechains exhibiting two or more physical-chemical properties can beincluded in multiple categories. For example, amino acid side chainshaving aromatic moieties that are further substituted with polarsubstituents, such as Tyr (Y), may exhibit both aromatic hydrophobicproperties and polar or hydrophilic properties, and can therefore beincluded in both the aromatic and polar categories. The appropriatecategorization of any amino acid will be apparent to those of skill inthe art, especially in light of the detailed disclosure provided herein.

Certain amino acid residues, called "helix breaking" amino acids, have apropensity to disrupt the structure of α-helices when contained atinternal positions within the helix. Amino acid residues exhibiting suchhelix-breaking properties are well-known in the art (see, e.g., Chou andFasman, Ann. Rev. Biochem. 47:251-276) and include Pro (P), Gly (G) andpotentially all D-amino acids (when contained in an L-peptide;conversely, L-amino acids disrupt helical structure when contained in aD-peptide). While these helix-breaking amino acid residues fall into thecategories defined above, these residues should not be used tosubstitute amino acid residues at internal positions within thehelix--they should only be used to substitute 1-3 amino acid residues atthe N-terminus and/or C-terminus of the peptide.

While the above-defined categories have been exemplified in terms of thegenetically encoded amino acids, the amino acid substitutions need notbe, and in certain embodiments preferably are not, restricted to thegenetically encoded amino acids. Indeed, many of the preferred peptidesof structure (I) contain genetically non-encoded amino acids. Thus, inaddition to the naturally occurring genetically encoded amino acids,amino acid residues in the core peptides of structure (I) may besubstituted with naturally occurring non-encoded amino acids andsynthetic amino acids.

Certain commonly encountered amino acids which provide usefulsubstitutions for the core peptides of structure (I) include, but arenot limited to, β-alanine (β-Ala) and other omega-amino acids such as3-aminopropionic acid, 2,3-diaminopropionic acid (Dpr), 4-aminobutyricacid and so forth; α-aminoisobutyric acid (Aib); ε-aminohexanoic acid(Aha); δ-aminovaleric acid (Ava); N-methylglycine or sarcosine (MeGly);ornithine (Orn); citrulline (Cit); t-butylalanine (t-BuA);t-butylglycine (t-BuG); N-methylisoleucine (MeIle); phenylglycine (Phg);cyclohexylalanine (Cha); norleucine (Nle); naphthylalanine (Nal);4-chlorophenylalanine (Phe(4-Cl)); 2-fluorophenylalanine (Phe(2-F));3-fluorophenylalanine (Phe(3-F)); 4-fluorophenylalanine (Phe(4-F));penicillamine (Pen); 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid(Tic); β-2-thienylalanine (Thi); methionine sulfoxide (MSO);homoarginine (hArg); N-acetyl lysine (AcLys); 2,4-diaminobutyric acid(Dbu); 2,3-diaminobutyric acid (Dab); p-aminophenylalanine (Phe(pNH₂));N-methyl valine (MeVal); homocysteine (hCys), homophenylalanine (hPhe)and homoserine (hSer); hydroxyproline (Hyp), homoproline (hPro),N-methylated amino acids and peptoids (N-substituted glycines).

The classifications of the genetically encoded and common non-encodedamino acids according to the categories defined above are summarized inTABLE III, below. It is to be understood that TABLE III is forillustrative purposes only and does not purport to be an exhaustive listof amino acid residues that can be used to substitute the core peptidesdescribed herein. Other amino acid residues not specifically mentionedherein can be readily categorized based on their observed physical andchemical properties in light of the definitions provided herein.

                  TABLE III                                                       ______________________________________                                        CLASSIFICATIONS OF COMMONLY ENCOUNTERED                                       AMINO ACIDS                                                                             Genetically   Non-Genetically                                       Classification                                                                          Encoded       Encoded                                               ______________________________________                                        Hydrophobic                                                                   Aromatic  F, Y, W       Phg, Nal, Thi, Tic, Phe (4-                                                   Cl), Phe (2-F), Phe (3-F),                                                    Phe (4-F), hPhe                                       Apolar    L, V, I, M, G, A, P                                                                         t-BuA, t-BuG, MeIle, Nle,                                                     MeVal, Cha, McGly, Aib                                Aliphatic A, V, L, I    b-Ala, Dpr, Aib, Aha, MeGly,                                                  t-BuA, t-BuG, MeIle, Cha,                                                     Nle, MeVal                                            Hydrophilic                                                                   Acidic    D, E                                                                Basic     H, K, R       Dpr, Orn, hArg, Phe (p-NH.sub.2),                                             Dbu, Dab                                              Polar     C, Q, N, S, T Cit, AcLys, MSO, bAla, hSer                           Helix-Breaking                                                                          P, G          D-Pro and other D-amino                                                       acids (in L-peptides)                                 ______________________________________                                    

While in most instances, the amino acids of the core peptides ofstructure (I) will be substituted with L-enantiomeric amino acids, thesubstitutions are not limited to L-enantiomeric amino acids. Thus, alsoincluded in the definition of "mutated" or "altered" forms are thosesituations where at least one L-amino acid is replaced with an identicalD-amino acid (e.g., L-Arg→D-Arg) or with a D-amino acid of the samecategory or subcategory (e.g., L-Arg→D-Lys), and vice versa. Indeed, incertain preferred embodiments that are suitable for oral administrationto animal subjects, the peptides may advantageously be composed of atleast one D-enantiomeric amino acid. Peptides containing such D-aminoacids are thought to be more stable to degradation in the oral cavity,gut or serum than are peptides composed exclusively of L-amino acids.

As noted above, D-amino acids tend to disrupt the structure of α-heliceswhen contained at internal positions of an α-helical L-peptide. As aconsequence, D-amino acids should not be used to substitute internalL-amino acids; D-amino acid substitutions should be limited to 1-3 aminoacid residues at the N-terminus and/or C-terminus of the peptide.Preferably, only the amino acid at the N- and/or C-terminus issubstituted with a D-amino acid.

Using the amino acid residue classifications described above inconjunction with the Schiffer-Edmundson helical wheel and helical netdiagram presentations of the core peptides of structure (I), as well asthe detailed description of the desired properties provided herein,altered or mutated forms of the core peptides of structure (I) thatsubstantially retain the amphipathic and other properties of the helix,and which are therefore considered to be within the scope of the presentinvention, can be readily obtained.

In a preferred embodiment of the invention, altered or mutated forms ofthe core peptides of structure (I) are obtained by fixing thehydrophilic or hydrophobic residues according to structure (I) andsubstituting at least one non-fixed residue with another amino acid,preferably conservatively, i.e., with another amino acid of the samecategory or sub-category. The residues composing the basic and/orhydrophobic clusters can also be fixed according to structure (I), andat least one non-fixed residue substituted, preferably conservatively.

In another preferred embodiment, altered or mutated forms of the corepeptides of structure (I) are obtained by fixing the hydrophilic aminoacid residues positioned within the hydrophilic face of the helixaccording to structure (I) and substituting at least one non-fixed aminoacid residue with another amino acid, preferably with another amino acidresidue of the same category or sub-category. Referring to FIG. 2A, itcan be seen that residues 1, 4, 7, 8, 11, 12, 14, 15 and 18 arepositioned within the hydrophilic face of the amphipathic helix formedby the core peptides of structure (I). Of these residues, all arehydrophilic except for residue 1, which may be either hydrophilic orhydrophobic. Thus, in one preferred embodiment, residues 4, 7, 8, 11,12, 14, 15 and 18 are fixed according to structure (I) and at least oneof residues 1, 2, 3, 5, 6, 9, 10, 13, 16 and 17 is substituted withanother amino acid of the same category, preferably with another aminoacid of the same sub-category. Alternatively, residue 1 is also fixedaccording to structure (I) and at least one of residues 2, 3, 5, 6, 9,10, 13, 16 and 17 is substituted.

In a particularly preferred embodiment, the C-terminal basic cluster(residues 14, 16 and 18) is also fixed according to structure (I), andonly residues 2, 3, 5, 6, 9, 10, 13 and/or 17 are substituted.

In another particularly preferred embodiment, the hydrophobic cluster(residues 3, 6, 9 and 10) is also fixed according to structure (I), andonly residues 2, 5, 13, 16 and/or 17 are substituted.

In still another particularly preferred embodiment, both the basic andhydrophobic clusters are fixed and only residues 2, 5, 13 and/or 17 aresubstituted.

In another preferred embodiment of the invention, altered or mutatedforms of the core peptides of the invention are obtained by fixing thehydrophobic amino acid residues positioned within the hydrophobic faceof the helix and substituting at least one non-fixed amino acid residuewith another amino acid residue, preferably with another residue of thesame category or sub-category.

Referring to FIG. 2A, it can be seen that residues 2, 3, 5, 6, 9, 10,13, 16 and 17 are positioned within the hydrophobic face. Of these, allare hydrophobic except for residue 16, which is hydrophilic. Thus, inone preferred embodiment residues 2, 3, 5, 6, 9, 10, 13 and 17 are fixedaccording to structure (I) and at least one of residues 1, 4, 7, 8, 11,12, 14, 15, 16 and 18 is substituted with another amino acid residue,preferably with another amino acid of the same category or subcategory.

In a particularly preferred embodiment, the C-terminal basic cluster(residues 14, 16 and 18) is also fixed, and only residues 1, 4, 7, 8,11, 12 and/or 15 are substituted.

In another embodiment, altered or mutated forms of the peptides ofstructure (I) are obtained by fixing all of the amino acid residuesresiding within the hydrophobic or hydrophilic face of the helix andsubstituting, preferably conservatively, at least one amino acid residueresiding in the other face with another amino acid residue. The residuescomprising the hydrophobic cluster and/or the basic cluster may also beoptionally fixed according to structure (I), as previously discussed.

In another embodiment of the invention, the altered or mutated forms ofstructure (I) are obtained by substituting at least one amino acid witha non-conservative amino acid. Those of skill in the art will recognizethat such substitutions should not substantially alter the amphipathicand/or structural properties of the helix discussed, supra. Thus, incertain instances it may be desirable to substitute one or more pairs ofamino acids so as to preserve the net properties of the helix. Furtherguidance for selecting appropriate amino acid substitutions is providedby the peptide sequences listed in TABLE IX (see, Section 8.3, infra).

In still another embodiment of the invention, the first one to fouramino acid residues at the N-terminus and/or C-terminus of the corepeptides of structure (I) are substituted with one or more amino acidresidues, or one or more peptide segments, that are known to conferstability to regions of α-helical secondary structure ("end-cap"residues or segments). Such end-cap residues and segments are well-knownin the art (see, e.g., Richardson and Richardson, 1988, Science240:1648-1652; Harper et al., 1993, Biochemistry 32(30):7605-7609;Dasgupta and Bell, 1993, Int. J. Peptide Protein Res. 41:499-511; Sealeet al., 1994, Protein Science 3:1741-1745; Doig et al., 1994,Biochemistry 33:3396-3403; Zhou et al., 1994, Proteins 18:1-7; Doig andBaldwin, 1995, Protein Science 4:1325-1336; Odaert et al., 1995,Biochemistry 34:12820-12829; Petrukhov et al., 1996, Biochemistry35:387-397; Doig et al., 1997, Protein Science 6:147-155).Alternatively, the first one to four N-terminal and/or C-terminal aminoacid residues of structure (I) can be replaced with peptidomimeticmoieties that mimic the structure and/or properties of end-cap residuesor segments. Suitable end-cap mimetics are well-known in the art, andare described, for example, in Richardson and Richardson, 1988, Science240:1648-1652; Harper et al., 1993, Biochemistry 32(30):7605-7609;Dasgupta and Bell, 1993, Int. J. Peptide Protein Res. 41:499-511; Sealeet al., 1994, Protein Science 3:1741-1745; Doig et al., 1994,Biochemistry 33:3396-3403; Zhou et al., 1994, Proteins 18:1-7; Doig andBaldwin, 1995, Protein Science 4:1325-1336; Odaert et al., 1995,Biochemistry 34:12820-12829; Petrukhov et al., 1996, Biochemistry35:387-397; Doig et al., 1997, Protein Science 6:147-155).

While structure (I) contains 18 specified amino acid residue positions,it is to be understood that the core peptides of the invention cancontain fewer than 18 amino acid residues. Indeed, truncated orinternally deleted forms of structure (I) containing as few as 14-15amino acid residues that substantially retain the overallcharacteristics and properties of the amphipathic helix formed by thecore peptides of structure (I) are considered to be within the scope ofthe present invention.

Truncated forms of the peptides of structure (I) are obtained bydeleting one or more amino acids from the N- and/or C-terminus ofstructure (I). Internally deleted forms of structure (I) are obtained bydeleting one or more amino acids from internal positions within thepeptide of structure (I). The internal amino acid residues deleted mayor may not be consecutive residues.

Those of skill in the art will recognize that deleting an internal aminoacid residue from a core peptide of structure (I) will cause the planeof the hydrophilic-hydrophobic interface of the helix to rotate by 100°at the point of the deletion. As such rotations can significantly alterthe amphipathic properties of the resultant helix, in a preferredembodiment of the invention amino acid residues are deleted so as tosubstantially retain the alignment of the plane of thehydrophilic-hydrophobic interface along the entire long axis of thehelix.

This can be conveniently achieved by deleting a sufficient number ofconsecutive or non-consecutive amino acid residues such that onecomplete helical turn is deleted. An idealized α-helix contains 3.6residues per turn. Thus, in a preferred embodiment, groups of 3-4consecutive or non-consecutive amino acid residues are deleted. Whether3 amino acids or 4 amino acids are deleted will depend upon the positionwithin the helix of the first residue to be deleted. Determining theappropriate number of consecutive or non-consecutive amino acid residuesthat constitute one complete helical turn from any particular startingpoint within an amphipathic helix is well within the capabilities ofthose of skill in the art.

Due to the surmised importance of the basic cluster at the C-terminus ofthe core peptides of structure (I) in stabilizing the helix and theimportance of the hydrophobic cluster in effecting lipid binding andLCAT activation, in preferred embodiments of the invention, residuescomprising the basic and hydrophobic clusters are not deleted. Thus, inpreferred embodiments, residues 14, 16 and 18 (basic cluster) andresidues 3, 6, 9 and 10 (hydrophobic cluster) are not deleted.

The core peptides of structure (I) can also be extended at one or bothtermini or internally with additional amino acid residues that do notsubstantially interfere with, and in some embodiments even enhance, thestructural and/or functional properties of the peptides. Indeed,extended core peptides containing as many as 19, 20, 21, 22 or even moreamino acid residues are considered to be within the scope of the presentinvention. Preferably, such extended peptides will substantially retainthe net amphipathicity and other properties of the peptides of structure(I). Of course, it will be recognized that adding amino acids internallywill rotate the plane of the hydrophobic-hydrophilic interface at thepoint of the insertion in a manner similar to that described above forinternal deletions. Thus, the considerations discussed above inconnection with internal deletions apply to internal additions, as well.

In one embodiment, the core peptides are extended at the N- and/orC-terminus by least one helical turn. Preferably, such extensions willstabilize the helical secondary structure in the presence of lipids,such as the end-cap amino acids and segments previously described.

In a particularly preferred embodiment, the core peptide of structure(I) is extended at the C-terminus by a single basic amino acid residue,preferably Lys (K).

Also included within the scope of the present invention are "blocked"forms of the ApoA-I agonist, i.e., forms of the ApoA-I agonists in whichthe N- and/or C-terminus is blocked with a moiety capable of reactingwith the N-terminal --NH₂ or C-terminal --C(O)OH. It has been discoveredthat removing the N- and/or C-terminal charges of the ApoA-I agonists ofthe invention containing 18 or fewer amino acid residues (bysynthesizing N-acylated peptide amides/ester/hydrazides/alcohols andsubstitutions thereof) results in agonists which approach, and in someembodiments even exceed, the activity of the unblocked form of theagonist. For example, while peptide 210 (SEQ ID NO:210) exhibits 46%LCAT activation as compared with native ApoA-I, an N- and C-terminalblocked form of this peptide, peptide 193 (SEQ ID NO:193), exhibits 96%LCAT activation in the same assay. In some embodiments containing 22amino acids, blocking the N- or C-terminus results in ApoA-I agonistswhich exhibit lower activity than the unblocked forms. However, blockingboth the N- and C-termini of ApoA-I agonists composed of 22 amino acidsis expected to restore activity. Thus, in a preferred embodiment of theinvention, either the N- and/or C-terminus (preferably both termini) ofcore peptides containing 18 or fewer amino acids are blocked, whereasthe N- and C-termini of peptides containing more than 18 amino acids areeither both blocked or both unblocked. Typical N-terminal blockinggroups include RC(O)--, where R is --H, (C₁ -C₆) alkyl, (C₁ -C₆)alkenyl, (C₁ -C₆) alkynyl, (C₅ -C₂₀) aryl, (C₆ -C₂₆) alkaryl, 5-20membered heteroaryl or 6-26 membered alkheteroaryl. Preferred N-terminalblocking groups include acetyl, formyl and dansyl. Typical C-terminalblocking groups include --C(O)NRR and --C(O)OR, where each R isindependently defined as above. Preferred C-terminal blocking groupsinclude those where each R is independently methyl. While not intendingto be bound by any particular theory, it is believed that such terminalblocking groups stabilize the α-helix in the presence of lipids (see,e.g., Venkatachelapathi et al., 1993, PROTEINS: Structure, Function andGenetics 15:349-359).

The native structure of ApoA-I contains eight helical units that arethought to act in concert to bind lipids (Nakagawa et al., 1985, J. Am.Chem. Soc. 107:7087-7092; Anantharamaiah et al., 1985, J. Biol. Chem.260:10248-10262; Vanloo et al., 1991, J. Lipid Res. 32:1253-1264; Mendezet al., 1994, J. Clin. Invest. 94:1698-1705; Palgunari et al., 1996,Arterioscler. Thromb. Vasc. Biol. 16:328-338; Demoor et al., 1996, Eur.J. Biochem. 239:74-84). Thus, also included in the present invention areApoA-I agonists comprised of dimers, trimers, tetramers and even higherorder polymers ("multimers") of the core peptides described herein. Suchmultimers may be in the form of tandem repeats, branched networks orcombinations thereof. The core peptides may be directly attached to oneanother or separated by one or more linkers.

The core peptides that comprise the multimers may be the peptides ofstructure (I), analogues of structure (I), mutated forms of structure(I), truncated or internally deleted forms of structure (I), extendedforms of structure (I) and/or combinations thereof. The core peptidescan be connected in a head-to-tail fashion (i.e., N-terminus toC-terminus), a head-to-head fashion, (i.e., N-terminus to N-terminus), atail-to-tail fashion (i.e., C-terminus to C-terminus), or combinationsthereof.

In one embodiment of the invention, the multimers are tandem repeats oftwo, three, four and up to about ten core peptides. Preferably, themultimers are tandem repeats of from 2 to 8 core peptides. Thus, in oneembodiment, the ApoA-I agonists of the invention comprise multimershaving the following structural formula: ##STR1## wherein: each m isindependently an integer from 0 to 1, preferably 1;

n is an integer from 0 to 10, preferably 0 to 8;

each "HH" independently represents a core peptide or peptide analogue ofstructure (I) or a mutated, truncated, internally deleted or extendedform thereof as described herein;

each "LL" independently represents a linker; and

each "-" independently designates a covalent linkage.

In structure (II), the linker LL can be any bifunctional moleculecapable of covalently linking two peptides to one another. Thus,suitable linkers are bifunctional molecules in which the functionalgroups are capable of being covalently attached to the N- and/orC-terminus of a peptide. Functional groups suitable for attachment tothe N- or C-terminus of peptides are well known in the art, as aresuitable chemistries for effecting such covalent bond formation.

The linker may be flexible, rigid or semi-rigid, depending on thedesired properties of the multimer. Suitable linkers include, forexample, amino acid residues such as Pro or Gly or peptide segmentscontaining from about 2 to about 5, 10, 15 or 20 or even more aminoacids, bifunctional organic compounds such as H₂ N(CH₂)_(n) COOH where nis an integer from 1 to 12, and the like. Examples of such linkers, aswell as methods of making such linkers and peptides incorporating suchlinkers are well-known in the art (see, e.g., Hunig et al., 1974, Chem.Ber. 100:3039-3044; Basak et al., 1994, Bioconjug. Chem. 5(4):301-305).

In a preferred embodiment of the invention, the tandem repeats areinternally punctuated by a single proline residue. To this end, in thoseinstances where the core peptides are terminated at their N- orC-terminus with proline, such as, e.g., where X₁ in structure (I) is Pro(P) or D-Pro (p), m in structure (II) is preferably 0. In thoseinstances where the core peptides do not contain an N- or C-terminalproline, LL is preferably Pro (P) or D-Pro (p) and m is preferably 1.

In certain embodiments of the invention, it may be desirable to employcleavable linkers that permit the release of one or more helicalsegments (HH) under certain conditions. Suitable cleavable linkersinclude peptides having amino acid sequences that are recognized byproteases, oligonucleotides that are cleaved by endonucleases andorganic compounds that can be cleaved via chemical means, such as underacidic, basic or other conditions. Preferably, the cleavage conditionswill be relatively mild so as not to denature or otherwise degrade thehelical segments and/or non-cleaved linkers composing the multimericApoA-I agonists.

Peptide and oligonucleotide linkers that can be selectively cleaved, aswell as means for cleaving the linkers are well known and will bereadily apparent to those of skill in the art. Suitable organic compoundlinkers that can be selectively cleaved will be apparent to those ofskill in the art, and include those described, for example, in WO94/08051, as well as the references cited therein.

In a preferred embodiment, the linkers employed are peptides that aresubstrates for endogenous circulatory enzymes, thereby permitting themultimeric ApoA-I agonists to be selectively cleaved in vivo. Endogenousenzymes suitable for cleaving the linkers include, for example,proapolipoprotein A-I propeptidase. Appropriate enzymes, as well aspeptide segments that act as substrates for such enzymes, are well-knownin the art (see, e.g., Edelstein et al., 1983, J. Biol. Chem.258:11430-11433; Zanis, 1983, Proc. Natl. Acad. Sci. U.S.A.80:2574-2578).

As discussed above, a key feature of the core peptides of the inventionis their ability to form intermolecular hydrogen-bonds or salt bridgeswhen arranged in an antiparallel fashion. Thus, in a preferredembodiment of the invention, linkers of sufficient length andflexibility are used so as to permit the helical segments (HH) ofstructure (II) to align in an antiparallel fashion and formintermolecular hydrogen-bonds or salt bridges in the presence of lipids.

Linkers of sufficient length and flexibility include, but are notlimited to, Pro (P), Gly (G), Cys--Cys, H₂ N--(CH₂)_(n) --C(O)OH where nis 1 to 12, preferably 4 to 6; H₂ N-aryl-C(O)OH and carbohydrates.

Alternatively, as the native apolipoproteins permit cooperative bindingbetween antiparallel helical segments, peptide linkers which correspondin primary sequence to the peptide segments connecting adjacent helicesof the native apolipoproteins, including, for example, ApoA-I, ApoA-II,ApoA-IV, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE and ApoJ can beconveniently used to link the core peptides. These sequences are wellknown in the art (see, e.g., Rosseneu et al., "Analysis of the Primaryand of the Secondary Structure of the Apolipoproteins," In: Structureand Function of Lipoproteins, Ch. 6, 159-183, CRC Press, Inc., 1992).

Other linkers which permit the formation of intermolecular hydrogenbonds or salt bridges between tandem repeats of antiparallel helicalsegments include peptide reverse turns such as β-turns and γ-turns, aswell as organic molecules that mimic the structures of peptide β-turnsand/or γ-turns. Generally, reverse turns are segments of peptide thatreverse the direction of the polypeptide chain so as to allow a singlepolypeptide chain to adopt regions of antiparallel β-sheet orantiparallel α-helical structure. β-turns generally are composed of fouramino acid residues and γ-turns are generally composed of three aminoacid residues.

The conformations and sequences of many peptide β-turns have beenwell-described in the art and include, by way of example and notlimitation, type-I, type-I', type-II, type-II', type-III, type-III',type-IV, type-V, type-V', type-VIa, type-VIb, type-VII and type-VIII(see, Richardson, 1981, Adv. Protein Chem. 34:167-339; Rose et al.,1985, Adv. Protein Chem. 37:1-109; Wilmot et al., 1988, J. Mol. Biol.203:221-232; Sibanda et al., 1989, J. Mol. Biol. 206:759-777; Tramontanoet al., 1989, Proteins: Struct. Funct. Genet. 6:382-394).

The specific conformations of short peptide turns such as β-turns dependprimarily on the positions of certain amino acid residues in the turn(usually Gly, Asn or Pro). Generally, the type-I β-turn is compatiblewith any amino acid residue at positions 1 through 4 of the turn, exceptthat Pro cannot occur at position 3. Gly predominates at position 4 andPro predominates at position 2 of both type-I and type-II turns. Asp,Asn, Ser and Cys residues frequently occur at position 1, where theirside chains often hydrogen-bond to the NH of residue 3.

In type-II turns, Gly and Asn occur most frequently at position 3, asthey adopt the required backbone angles most easily. Ideally, type-I'turns have Gly at positions 2 and 3, and type-II' turns have Gly atposition 2. Type-III turns generally can have most amino acid residues,but type-III' turns usually require Gly at positions 2 and 3. Type-VIaand VIb turns generally have a cis peptide bond and Pro as an internalresidue. For a review of the different types and sequences of β-turns inproteins and peptides the reader is referred to Wilmot et al., 1988, J.Mol. Biol. 203:221-232.

The conformation and sequences of many peptide γ-turns have also beenwell-described in the art (see, e.g., Rose et al., 1985, Adv. ProteinChem. 37:1-109; Wilmer-White et al., 1987, Trends Biochem. Sci.12:189-192; Wilmot et al., 1988, J. Mol. Biol. 203:221-232; Sibanda etal., 1989, J. Mol. Biol. 206:759-777; Tramontano et al., 1989, Proteins:Struct. Funct. Genet. 6:382-394). All of these types of β-turns andγ-turn structures and their corresponding sequences, as well as laterdiscovered peptide β-turns and γ-turn structures and sequences, arespecifically contemplated by the invention.

Alternatively, the linker (LL) may comprise an organic molecule ormoiety that mimics the structure of a peptide β-turn or γ-turn. Suchβ-turn and/or γ-turn mimetic moieties, as well as methods forsynthesizing peptides containing such moieties, are well known in theart, and include, among others, those described in Giannis and Kolter,1993 Angew. Chem. Intl. Ed. Eng. 32:1244-1267; Kahn et al., 1988, J.Molecular Recognition 1:75-79; and Kahn et al., 1987, Tetrahedron Lett.28:1623-1626.

In still another embodiment of the invention, the multimers are in theform of branched networks (see, e.g., FIG. 7). Such networks areconveniently obtained through the use of multifunction linking moietiesthat permit more than two helical units to be attached to a simplelinking moiety. Thus, branched networks employ molecules having three,four or even more functional groups that are capable of covalentlyattaching to the N- and/or C-terminus of a peptide. Suitable linkingmoieties include, for example, amino acid residues having side chainsbearing hydroxyl, sulfanyl, amino, carboxyl, amide and/or esterfunctionalities, such as, for example, Ser (S), Thr (T), Cys (C), Tyr(Y), Asn (N), Gln (Q), Lys (K), Arg (R), Orn, Asp (D) and Glu (E); orother organic molecules containing such functional groups.

The helical segments attached to a single linking moiety need not beattached via like termini. Indeed, in some embodiments the helicalsegments are attached to a single linking moiety so as to be arranged inan antiparallel fashion, i.e., some of the helices are attached viatheir N-termini, others via their C-termini.

The helical segments can be attached directly to the linking moiety, ormay be spaced from the linking moiety by way of one or more bifunctionallinkers (LL), as previously described.

Referring to FIGS. 7A and 7B, it can be seen that a branched network canbe described in terms of the number of "nodes" comprising the network,where each multifunctional linking moiety constitutes a node. In FIGS.7A and 7B, helical segments (i.e., core peptides of the invention) areillustrated as cylinders, and multifunctional linking moieties (ornodes) as circles (), where the number of lines emanating from thecircle indicates the "order" (or number of functional groups) of themultifunctional linking moiety.

The number of nodes in the network will generally depend on the totaldesired number of helical segments, and will typically be from about 1to 2. Of course, it will be appreciated that for a given number ofdesired helical segments, networks having higher order linking moietieswill have fewer nodes. For example, referring to FIGS. 7A and 7B, atertiary-order network (i.e., a network having trifunctional linkingmoieties) of seven helical units has three nodes (FIG. 7A), whereas aquaternary order network (i.e., a network having tetrafunctional linkingmoieties) of seven helical units has only two nodes (FIG. 7B).

The networks may be of uniform order, i.e., networks in which all nodesare, for example, trifunctional or tetrafunctional linking moieties, ormay be of mixed order, e.g., networks in which the nodes are mixturesof, for example, trifunctional and tetrafunctional linking moieties. Ofcourse, it is to be understood that even in uniform order networks thelinking moieties need not be identical. A tertiary order network mayemploy, for example, two, three, four or even more differenttrifunctional linking moieties.

Like the linear multimers, the helical segments comprising the branchednetwork may be, but need not be, identical.

An example of such a mixed order branched network is illustrated in FIG.7C. In FIG. 7C, helical segments (i.e., core peptides of the invention)are illustrated as cylinders and multifunctional linking moieties ascircles (), where the number of lines emanating from the circleindicates the "order" (or number of functional groups) of themultifunctional linking moiety. Lines connecting helical segmentsrepresent bifunctional linkers LL, as previously described. Helicalsegments which comprise the branched networks may be tandem repeats ofcore peptides, as previously described.

In one illustrative embodiment, the branched networks of the inventionare described by the formula: ##STR2## wherein: each X is independently##STR3## each HH is independently a core peptide of structure (I) or ananalogue or mutated, truncated, internally deleted or extended formthereof as described herein;

each LL is independently a bifunctional linker;

each m is independently an integer from 0 to 1;

each n is independently an integer from 0 to 8;

N_(ya) and N_(yb) are each independently a multifunctional linkingmoiety where y_(a) and y_(b) represent the number of functional groupson N_(ya) and N_(yb), respectively;

each y_(a) or y_(b) is independently an integer from 3 to 8;

p is an integer from 0 to 7; and

each "-" independently designates a covalent bond.

In a preferred embodiment, the branched network comprises a "Lys-tree,"i.e., a network wherein the multifunctional linking moiety is one ormore Lys (K) residues (see, e.g., FIG. 7D).

In one illustrative embodiment, the "Lys tree" branched networks of theinvention are described by the formulae: ##STR4## wherein: each X isindependently ##STR5## each HH is independently a core peptide orpeptide analogue of structure (I) or a mutated, truncated, internallydeleted or extended form thereof as described herein;

each LL is independently a bifunctional linker;

each n is independently an integer from 0 to 8;

each m is independently an integer from 0 to 1;

R₁ is --OR or --NRR; and

each R is independently --H, (C₁ -C₆) alkyl, (C₁ -C₆) alkenyl, (C₁ -C₆)alkynyl; (C₁ -C₂₀) aryl (C₆ -C₂₆) alkaryl, alkaryl, 5-20 memberedheteroaryl or 6-26 membered alkheteroaryl.

5.1.1. Analysis of Structure and Function

The structure and function of the core peptides or peptide analogues ofthe invention, as well as ApoA-I agonists composed of such corepeptides, including the multimeric forms described above, can be assayedin order to select active agonists or mimetics of ApoA-I. For example,the core peptides or peptide analogues can be assayed for their abilityto form α-helices in the presence of lipids, to bind lipids, to formcomplexes with lipids, to activate LCAT, to promote cholesterol efflux,etc.

Methods and assays for analyzing the structure and/or function of thepeptides are well-known in the art. Preferred methods are provided inthe working examples, infra. For example, the circular dichroism (CD)and nuclear magnetic resonance (NMR) assays described in Section 7,infra, can be used to analyze the structure of the peptides or peptideanalogues--particularly the degree of helicity in the presence oflipids. The ability to bind lipids can be determined using thefluorescence spectroscopy assay described in Section 7, infra. Theability of the peptides and/or peptide analogues to activate LCAT can bereadily determined using the LCAT activation described in Section 8,infra. The in vitro and in vivo assays described in Section 9, 10 and11, infra, can be used to evaluate the half-life, distribution,cholesterol efflux and effects on RCT.

Generally, core peptides and/or peptide analogues according to theinvention which exhibit the properties listed in TABLE IV, infra, areconsidered to be active.

                  TABLE IV                                                        ______________________________________                                        PROPERTIES OF ACTIVE PEPTIDES                                                                              PREFERRED                                        PROPERTY          RANGE      RANGE                                            ______________________________________                                        % Helicity in the presence of                                                                   ≧60%                                                                              ≧80%                                      lipids (Ri = 30) (unblocked 22-                                               amino acid residue peptides)                                                  % Helicity in the presence of                                                                   ≧40%                                                                              ≧60%                                      lipids (Ri = 30) (unblocked 18-                                               amino acid residue peptides)                                                  % Helicity in the presence of                                                                   ≧60%                                                                              ≧80%                                      lipids (Ri = 30) (blocked 18-amino                                            acid residue peptides and shorter                                             peptides)                                                                     Lipid Binding (in the presence of                                                               0.5-10 μM                                                SUVs)             peptide                                                                       R.sub.i = 1-50                                              LCAT activation   ≧38%                                                                              ≧80%                                      ______________________________________                                    

R_(i) is lipid:peptide molar ratio.

As illustrated in the working examples, infra, core peptides whichexhibit a high degree of LCAT activation (≧38%) generally possesssignificant α-helical structure in the presence of lipidic smallunilamellar vesicles (SUVs) (≧60% helical structure in the case ofunblocked peptides containing 22 or more amino acid residues and blockedpeptides containing 18 or fewer amino acid residues; ≧40% helicalstructure in the case of unblocked peptides containing 18 or fewer aminoacids), and those peptides which exhibit little or no LCAT activationpossess little α-helical structure. However, in certain instances,peptides which exhibit significant helical structure in the presence oflipids do not effect significant LCAT.

Similarly, while core peptides that exhibit significant LCAT activationtypically bind lipids, in certain instances peptides which exhibit lipidbinding do not effect significant LCAT activation.

As a consequence, it will be recognized by those of skill in the artthat while the ability of the core peptides described herein to formα-helices (in the presence of lipids) and to bind lipids is critical foractivity, in many instances these properties may not be sufficient.Thus, in a preferred embodiment core peptides of the invention aresubjected to a series of screens to select for core peptides exhibitingsignificant pharmacological activity.

In a first step, a core peptide is screened for its ability to form anα-helix in the presence of lipids using the CD assay described inSection 7, infra. Those peptides which are at least 40% helical(unblocked peptides containing 18 or fewer amino acids) or 60% helical(blocked peptides containing 18 or fewer amino acids; unblocked peptidescontaining 22 or more amino acids) in the presence of lipids (at a conc.of about 5 μM and a lipid:peptide molar ratio of about 30) are thenscreened for their ability to bind lipids using the fluorescence assaydescribed in Section 7, infra. Of course, only those core peptides whichcontain a fluorescent Trp (W) or Nal residue are screened for lipidbinding via fluorescence. However, for peptides which do not containfluorescent residues, binding to lipids is obvious when helicityincreases in the presence of lipids.

Core peptides which exhibit lipid binding in the presence of SUVs(0.5-10 μM peptide; lipid:peptide molar ratio in the range of 1 to 50)are then screened for pharmacological activity. Of course, thepharmacological activity screened for will depend upon the desired useof the ApoA-I agonists. In a preferred embodiment, the core peptides arescreened for their ability to activate LCAT, as peptides which activateLCAT are particularly useful in the methods described herein. Corepeptides which exhibit at least about 38% LCAT activation as comparedwith native human ApoA-I (as determined using the LCAT activation assaydescribed in Section 8, infra), are preferred, with core peptidesexhibiting 50%, 60%, 70%, 80% or even 90% or more being particularlypreferred.

5.1.2. Preferred Embodiments

The ApoA-I agonists of the invention can be further defined by way ofpreferred embodiments.

In one preferred embodiment, the ApoA-I agonists are 18 amino acidresidue peptides according to structure (I), or the N-terminal acylatedand/or C-terminal amidated or esterified forms thereof.

In another preferred embodiment, the ApoA-I agonists are 18 amino acidresidue peptides according to structure (I), or the N-terminal acylatedand/or C-terminal amidated or esterified forms thereof, in which:

X₂ is Ala (A), Val (V) or Leu (L);

X₄ is Asp (D) or Glu (E);

X₇ is Arg (R), Lys (K) or orn;

X₈ is Asp (D) or Glu (E);

X₁₁ is Glu (E) or Asn (N);

X₁₂ is Glu (E);

X₁₄ is Arg (R), Lys (K), Orn or Leu (L);

X₁₆ is Arg (R), Lys (K) or Orn; and/or

X₁₈ is Arg (R), Lys (K) or Orn, and

X₁, X₃, X₅, X₆, X₉, X₁₀, X₁₃, X₁₅ and X₁₇ are as previously defined forstructure (I).

In another preferred embodiment, the ApoA-I agonists are 18 amino acidresidue peptides according to structure (I), or the N-terminal acylatedand/or C-terminal amidated or esterified forms thereof, in which whenX₁₁ is Asn (N), X₁₄ is Leu (L) and when X₁₁ is other than Asn (N), X₁₄is other than Leu (L). Particularly preferred embodiments according tothis aspect of the invention are those peptides, or the N-terminalacylated and/or C-terminal amidated or esterified forms thereof, inwhich the various X_(n) in structure (I) are defined as in the precedingparagraph. An exemplary particularly preferred embodiment according tothis aspect of the invention is the peptide 209 (PVLDLFRELLNELLQKLK; SEQID NO:209), and the N-terminal acylated and/or C-terminal amidated oresterified forms thereof.

In still another preferred embodiment, the ApoA-I agonists are alteredor mutated forms of the peptides according to structure (I), or theN-terminal acylated and/or C-terminal amidated or esterified formsthereof, in which

X₁ is other than Asp (D);

X₉ is other than Gly (G);

X₁₀ is other than Gly (G);

X₁₂ is other than Leu (L); and

X₁₃ is other than Gly (G).

In still another preferred embodiment, the ApoA-I agonists are selectedfrom the group of peptides set forth below:

peptide 191 PVLDLLRELLEELKQKLK* (SEQ ID NO:191);

peptide 192 PVLDLFKELLEELKQKLK* (SEQ ID NO:192);

peptide 193 PVLDLFRELLEELKQKLK* (SEQ ID NO:193);

peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194);

peptide 195 PVLELFKELLEELKQKLK* (SEQ ID NO:195);

peptide 196 PVLDLFRELLEELKNKLK* (SEQ ID NO:196);

peptide 197 PLLDLFRELLEELKQKLK* (SEQ ID NO:197);

peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198);

peptide 199 PVLDLFRELWEELKQKLK* (SEQ ID NO:199);

peptide 200 NVLDLFRELLEELKQKLK* (SEQ ID NO:200);

peptide 201 PLLDLFKELLEELKQKLK* (SEQ ID NO:201);

peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202);

peptide 203 AVLDLFRELLEELKQKLK* (SEQ ID NO:203);

peptide 204 PVLDFFRELLEELKQKLK* (SEQ ID NO:204);

peptide 205 PVLDLFREWLEELKQKLK* (SEQ ID NO:205);

peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206);

peptide 207 PVLELLKELLEELKQKLK* (SEQ ID NO:207);

peptide 208 PALELFKDLLEELRQRLK* (SEQ ID NO:208);

peptide 209 PVLDLFRELLNELLQKLK (SEQ ID NO:209);

peptide 210 PVLDLFRELLEELKQKLK (SEQ ID NO:210);

peptide 211 PVLDLFRELLEELOQOLO* (SEQ ID NO:211);

peptide 212 PVLDLFOELLEELOQOLK* (SEQ ID NO:212);

peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213);

peptide 214 pVLDLFRELLEELKQKLK* (SEQ ID NO:214);

peptide 215 PVLDLFRELLEEWKQKLK* (SEQ ID NO:215);

peptide 230 PVLELFERLLEDLQKKLK (SEQ ID NO:230);

peptide 231 PVLDLFRELLEKLEQKLK (SEQ ID NO:231);

peptide 232 PLLELFKELLEELKQKLK* (SEQ ID NO:232);

in either the N- and/or C-terminal blocked or unblocked forms.

In still another preferred embodiment, the ApoA-I agonists are selectedfrom the group of peptides set forth below:

peptide 191 PVLDLLRELLEELKQKLK* (SEQ ID NO:191);

peptide 192 PVLDLFKELLEELKQKLK* (SEQ ID NO:192);

peptide 193 PVLDLFRELLEELKQKLK* (SEQ ID NO:193);

peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194);

peptide 195 PVLELFKELLEELKQKLK* (SEQ ID NO:195);

peptide 196 PVLDLFRELLEELKNKLK* (SEQ ID NO:196);

peptide 197 PLLDLFRELLEELKQKLK* (SEQ ID NO:197);

peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198);

peptide 199 PVLDLFRELWEELKQKLK* (SEQ ID NO:199);

peptide 200 NVLDLFRELLEELKQKLK* (SEQ ID NO:200);

peptide 201 PLLDLFKELLEELKQKLK* (SEQ ID NO:201);

peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202);

peptide 203 AVLDLFRELLEELKQKLK* (SEQ ID NO:203);

peptide 204 PVLDFFRELLEELKQKLK* (SEQ ID NO:204);

peptide 205 PVLDLFREWLEELKQKLK* (SEQ ID NO:205);

peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206);

peptide 207 PVLELLKELLEELKQKLK* (SEQ ID NO:207);

peptide 208 PALELFKDLLEELRQRLK* (SEQ ID NO:208);

peptide 209 PVLDLFRELLNELLQKLK (SEQ ID NO:209);

peptide 210 PVLDLFRELLEELKQKLK (SEQ ID NO:210);

peptide 211 PVLDLFRELLEELOQOLO* (SEQ ID NO:211);

peptide 212 PVLDLFOELLEELOQOLK* (SEQ ID NO:212);

peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213);

peptide 214 pVLDLFRELLEELKQKLK* (SEQ ID NO:214);

peptide 215 PVLDLFRELLEEWKQKLK* (SEQ ID NO:215);

in either the N- and/or C-terminal blocked or unblocked forms.

In yet another preferred embodiment, the ApoA-I agonists are multimericforms according to structures II, III and/or IV in which each HH isindependently a peptide according to structure (I) or an N-terminalacylated and/or C-terminal amidated or esterified form thereof, or anyof the preferred peptides according to structure (I) described herein.

In yet another preferred embodiment, the core peptides that compose theApoA-I agonists are not any of the following peptides:

peptide 75: PVLDEFREKLNEELEALKQKLK (SEQ ID NO:75);

peptide 94: PVLDEFREKLNEALEALKQKLK (SEQ ID NO:94);

peptide 109: PVLDEFREKLNERLEALKQKLK (SEQ ID NO:109);

peptide 237: LDDLLQKWAEAFNQLLKK (SEQ ID NO:237);

peptide 238: EWLKAFYEKVLEKLKELF* (SEQ ID NO:238);

peptide 241: DWFKAFYDKVFEKFKEFF (SEQ ID NO:241);

peptide 242: GIKKFLGSIWKFIKAFVG (SEQ ID NO:242);

peptide 243: DWFKAFYDKVAEKFKEAF (SEQ ID NO:243);

peptide 244: DWLKAFYDKVAEKLKEAF (SEQ ID NO:244);

peptide 245: DWLKAFYDKVFEKFKEFF (SEQ ID NO:245);

peptide 246: EWLEAFYKKVLEKLKELF (SEQ ID NO:246);

peptide 247: DWFKAFYDKFFEKFKEFF (SEQ ID NO:247);

peptide 248: EWLKAFYEKVLEKLKELF (SEQ ID NO:248);

peptide 249: EWLKAEYEKVEEKLKELF* (SEQ ID NO:249);

peptide 250: EWLKAEYEKVLEKLKELF* (SEQ ID NO:250); and

peptide 251: EWLKAFYKKVLEKLKELF* (SEQ ID NO:251).

In a final preferred embodiment, the ApoA-I agonists are not any of thepeptides listed in TABLE IX (Section 8.3, infra) which exhibit an LCATactivation activity of less than 38% as compared with native humanApoA-I. 5.2 Synthesis and Purification of the ApoA-I Peptide Agonists

The core peptides of the invention may be prepared using virtually anyart-known technique for the preparation of peptides. For example, thepeptides may be prepared using conventional step-wise solution or solidphase peptide syntheses, or recombinant DNA techniques.

5.2.1 Chemical Synthesis

Core peptides may be prepared using conventional step-wise solution orsolid phase synthesis (see, e.g., Chemical Approaches to the Synthesisof Peptides and Proteins, Williams et al., Eds., 1997, CRC Press, BocaRaton Fla., and references cited therein; Solid Phase Peptide Synthesis:A Practical Approach, Atherton & Sheppard, Eds., 1989, IRL Press,Oxford, England, and references cited therein).

Alternatively, the peptides of the invention may be prepared by way ofsegment condensation, as described, for example, in Liu et al., 1996,Tetrahedron Lett. 37(7):933-936; Baca, et al., 1995, J. Am. Chem. Soc.117:1881-1887; Tam et al., 1995, Int. J. Peptide Protein Res.45:209-216; Schnolzer and Kent, 1992, Science 256:221-225; Liu and Tam,1994, J. Am. Chem. Soc. 116(10):4149-4153; Liu and Tam, 1994, Proc.Natl. Acad. Sci. U.S.A. 91:6584-6588; Yamashiro and Li, 1988, Int. J.Peptide Protein Res. 31:322-334). Segment condensation is a particularlyuseful method for synthesizing embodiments containing internal glycineresidues. Other methods useful for synthesizing the peptides of theinvention are described in Nakagawa et al., 1985, J. Am. Chem. Soc.107:7087-7092.

ApoA-I agonists containing N- and/or C-terminal blocking groups can beprepared using standard techniques of organic chemistry. For example,methods for acylating the N-terminus of a peptide or amidating oresterifying the C-terminus of a peptide are well-known in the art. Modesof carrying other modifications at the N- and/or C-terminus will beapparent to those of skill in the art, as will modes of protecting anyside-chain functionalities as may be necessary to attach terminalblocking groups.

Pharmaceutically acceptable salts (counter ions) can be convenientlyprepared by ion-exchange chromatography or other methods as are wellknown in the art.

Compounds of the invention which are in the form of tandem multimers canbe conveniently synthesized by adding the linker(s) to the peptide chainat the appropriate step in the synthesis. Alternatively, the helicalsegments can be synthesized and each segment reacted with the linker. Ofcourse, the actual method of synthesis will depend on the composition ofthe linker. Suitable protecting schemes and chemistries are well known,and will be apparent to those of skill in the art.

Compounds of the invention which are in the form of branched networkscan be conveniently synthesized using the trimeric and tetrameric resinsand chemistries described in Tam, 1988, Proc. Natl. Acad. Sci. U.S.A.85:5409-5413 and Demoor et al., 1996, Eur. J. Biochem. 239:74-84.Modifying the synthetic resins and strategies to synthesize branchednetworks of higher or lower order, or which contain combinations ofdifferent core peptide helical segments, is well within the capabilitiesof those of skill in the art of peptide chemistry and/or organicchemistry.

Formation of disulfide linkages, if desired, is generally conducted inthe presence of mild oxidizing agents. Chemical oxidizing agents may beused, or the compounds may simply be exposed to atmospheric oxygen toeffect these linkages. Various methods are known in the art, includingthose described, for example, by Tam et al., 1979, Synthesis 955-957;Stewart et al., 1984, Solid Phase Peptide Synthesis, 2d Ed., PierceChemical Company Rockford, Ill.; Ahmed et al., 1975, J. Biol. Chem.250:8477-8482; and Pennington et al., 1991 Peptides 1990 164-166, Giraltand Andreu, Eds., ESCOM Leiden, The Netherlands. An additionalalternative is described by Kamber et al., 1980, Helv. Chim. Acta63:899-915. A method conducted on solid supports is described byAlbericio, 1985, Int. J. Peptide Protein Res. 26:92-97. Any of thesemethods may be used to form disulfide linkages in the peptides of theinvention.

5.2.2 Recombinant Synthesis

If the peptide is composed entirely of gene-encoded amino acids, or aportion of it is so composed, the peptide or the relevant portion mayalso be synthesized using conventional recombinant genetic engineeringtechniques.

For recombinant production, a polynucleotide sequence encoding thepeptide is inserted into an appropriate expression vehicle, i.e., avector which contains the necessary elements for the transcription andtranslation of the inserted coding sequence, or in the case of an RNAviral vector, the necessary elements for replication and translation.The expression vehicle is then transfected into a suitable target cellwhich will express the peptide. Depending on the expression system used,the expressed peptide is then isolated by procedures well-established inthe art. Methods for recombinant protein and peptide production are wellknown in the art (see, e.g., Sambrook et al., 1989, Molecular Cloning ALaboratory Manual, Cold Spring Harbor Laboratory, New York; and Ausubelet al., 1989, Current Protocols in Molecular Biology, Greene PublishingAssociates and Wiley Interscience, New York each of which isincorporated by reference herein in its entirety.)

To increase efficiency of production, the polynucleotide can be designedto encode multiple units of the peptide separated by enzymatic cleavagesites--either homopolymers (repeating peptide units) or heteropolymers(different peptides strung together) can be engineered in this way. Theresulting polypeptide can be cleaved (e.g., by treatment with theappropriate enzyme) in order to recover the peptide units. This canincrease the yield of peptides driven by a single promoter. In apreferred embodiment, a polycistronic polynucleotide can be designed sothat a single mRNA is transcribed which encodes multiple peptides (i.e.,homopolymers or heteropolymers) each coding region operatively linked toa cap-independent translation control sequence; e.g., an internalribosome entry site (IRES). When used in appropriate viral expressionsystems, the translation of each peptide encoded by the mRNA is directedinternally in the transcript; e.g., by the IRES. Thus, the polycistronicconstruct directs the transcription of a single, large polycistronicmRNA which, in turn, directs the translation of multiple, individualpeptides. This approach eliminates the production and enzymaticprocessing of polyproteins and may significantly increase yield ofpeptide driven by a single promoter.

A variety of host-expression vector systems may be utilized to expressthe peptides described herein. These include, but are not limited to,microorganisms such as bacteria transformed with recombinantbacteriophage DNA or plasmid DNA expression vectors containing anappropriate coding sequence; yeast or filamentous fungi transformed withrecombinant yeast or fungi expression vectors containing an appropriatecoding sequence; insect cell systems infected with recombinant virusexpression vectors (e.g., baculovirus) containing an appropriate codingsequence; plant cell systems infected with recombinant virus expressionvectors (e.g., cauliflower mosaic virus or tobacco mosaic virus) ortransformed with recombinant plasmid expression vectors (e.g., Tiplasmid) containing an appropriate coding sequence; or animal cellsystems.

The expression elements of the expression systems vary in their strengthand specificities. Depending on the host/vector system utilized, any ofa number of suitable transcription and translation elements, includingconstitutive and inducible promoters, may be used in the expressionvector. For example, when cloning in bacterial systems, induciblepromoters such as pL of bacteriophage λ, plac, ptrp, ptac (ptrp-lachybrid promoter) and the like may be used; when cloning in insect cellsystems, promoters such as the baculovirus polyhedron promoter may beused; when cloning in plant cell systems, promoters derived from thegenome of plant cells (e.g., heat shock promoters; the promoter for thesmall subunit of RUBISCO; the promoter for the chlorophyll a/b bindingprotein) or from plant viruses (e.g., the 35S RNA promoter of CaMV; thecoat protein promoter of TMV) may be used; when cloning in mammaliancell systems, promoters derived from the genome of mammalian cells(e.g., metallothionein promoter) or from mammalian viruses (e.g., theadenovirus late promoter; the vaccinia virus 7.5K promoter) may be used;when generating cell lines that contain multiple copies of expressionproduct, SV40-, BPV- and EBV-based vectors may be used with anappropriate selectable marker.

In cases where plant expression vectors are used, the expression ofsequences encoding the peptides of the invention may be driven by any ofa number of promoters. For example, viral promoters such as the 35S RNAand 19S RNA promoters of CaMV (Brisson et al., 1984, Nature310:511-514), or the coat protein promoter of TMV (Takamatsu et al.,1987, EMBO J. 6:307-311) may be used; alternatively, plant promoterssuch as the small subunit of RUBISCO (Coruzzi et al., 1984, EMBO J.3:1671-1680; Broglie et al., 1984, Science 224:838-843) or heat shockpromoters, e.g., soybean hsp17.5-E or hsp17.3-B (Gurley et al., 1986,Mol. Cell. Biol. 6:559-565) may be used. These constructs can beintroduced into plant cells using Ti plasmids, Ri plasmids, plant virusvectors, direct DNA transformation, microinjection, electroporation,etc. For reviews of such techniques see, e.g., Weissbach & Weissbach,1988, Methods for Plant Molecular Biology, Academic Press, New York,Section VIII, pp. 421-463; and Grierson & Corey, 1988, Plant MolecularBiology, 2d Ed., Blackie, London, Ch. 7-9.

In one insect expression system that may be used to produce the peptidesof the invention, Autographa californica, nuclear polyhidrosis virus(AcNPV) is used as a vector to express the foreign genes. The virusgrows in Spodoptera frugiperda cells. A coding sequence may be clonedinto non-essential regions (for example the polyhedron gene) of thevirus and placed under control of an AcNPV promoter (for example, thepolyhedron promoter). Successful insertion of a coding sequence willresult in inactivation of the polyhedron gene and production ofnon-occluded recombinant virus (i.e., virus lacking the proteinaceouscoat coded for by the polyhedron gene). These recombinant viruses arethen used to infect Spodoptera frugiperda cells in which the insertedgene is expressed (e.g., see Smith et al., 1983, J. Virol. 46:584;Smith, U.S. Pat. No. 4,215,051). Further examples of this expressionsystem may be found in Current Protocols in Molecular Biology, Vol. 2,Ausubel et al., eds., Greene Publish. Assoc. & Wiley Interscience.

In mammalian host cells, a number of viral based expression systems maybe utilized. In cases where an adenovirus is used as an expressionvector, a coding sequence may be ligated to an adenovirustranscription/translation control complex, e.g., the late promoter andtripartite leader sequence. This chimeric gene may then be inserted inthe adenovirus genome by in vitro or in vivo recombination. Insertion ina non-essential region of the viral genome (e.g., region E1 or E3) willresult in a recombinant virus that is viable and capable of expressingpeptide in infected hosts. (e.g., See Logan & Shenk, 1984, Proc. Natl.Acad. Sci. (U.S.A.) 81:3655-3659). Alternatively, the vaccinia 7.5Kpromoter may be used, (see, e.g., Mackett et al., 1982, Proc. Natl.Acad. Sci. (U.S.A.) 79:7415-7419; Mackett et al., 1984, J. Virol.49:857-864; Panicali et al., 1982, Proc. Natl. Acad. Sci. 79:4927-4931).

Other expression systems for producing the peptides of the inventionwill be apparent to those having skill in the art.

5.2.3 Purification of Peptides

The peptides of the invention can be purified by art-known techniquessuch as reverse phase chromatography high performance liquidchromatography, ion exchange chromatography, gel electrophoresis,affinity chromatography and the like. The actual conditions used topurify a particular peptide will depend, in part, on synthesis strategyand on factors such as net charge, hydrophobicity, hydrophilicity, etc.,and will be apparent to those having skill in the art. Multimericbranched peptides can be purified, e.g., by ion exchange or sizeexclusion chromatography.

For affinity chromatography purification, any antibody whichspecifically binds the peptide may be used. For the production ofantibodies, various host animals, including but not limited to rabbits,mice, rats, etc., may be immunized by injection with a peptide. Thepeptide may be attached to a suitable carrier, such as BSA, by means ofa side chain functional group or linkers attached to a side chainfunctional group. Various adjuvants may be used to increase theimmunological response, depending on the host species, including but notlimited to Freund's (complete and incomplete), mineral gels such asaluminum hydroxide, surface active substances such as lysolecithin,pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpethemocyanin, dinitrophenol, and potentially useful human adjuvants suchas BCG (bacilli Calmette-Guerin) and Corynebacterium parvum.

Monoclonal antibodies to a peptide may be prepared using any techniquewhich provides for the production of antibody molecules by continuouscell lines in culture. These include but are not limited to thehybridoma technique originally described by Kohler and Milstein, 1975,Nature 256:495-497, or Kaprowski, U.S. Pat. No. 4,376,110 which isincorporated by reference herein; the human B-cell hybridoma technique)Kosbor et al., 1983, Immunology Today 4:72; Cote et al., 1983, Proc.Natl. Acad. Sci. U.S.A. 80:2026-2030); and the EBV-hybridoma technique(Cole et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R.Liss, Inc., pp. 77-96 (1985)). In addition, techniques developed for theproduction of "chimeric antibodies" Morrison et al., 1984, Proc. Natl.Acad. Sci. U.S.A. 81:6851-6855; Neuberger et al., 1984, Nature312:604-608; Takeda et al., 1985, Nature 314:452-454, Boss, U.S. Pat.No. 4,816,397; Cabilly, U.S. Pat. No. 4,816,567; which are incorporatedby reference herein) by splicing the genes from a mouse antibodymolecule of appropriate antigen specificity together with genes from ahuman antibody molecule of appropriate biological activity can be used.Or "humanized" antibodies can be prepared (see, e.g., Queen, U.S. Pat.No. 5,585,089 which is incorporated by reference herein). Alternatively,techniques described for the production of single chain antibodies (U.S.Pat. No. 4,946,778) can be adapted to produce peptide-specific singlechain antibodies.

Antibody fragments which contain deletions of specific binding sites maybe generated by known techniques. For example, such fragments includebut are not limited to F(ab')₂ fragments, which can be produced bypepsin digestion of the antibody molecule and Fab fragments, which canbe generated by reducing the disulfide bridges of the F(ab')₂ fragments.Alternatively, Fab expression libraries may be constructed (Huse et al.,1989, Science 246:1275-1281) to allow rapid and easy identification ofmonoclonal Fab fragments with the desired specificity for the peptide ofinterest.

The antibody or antibody fragment specific for the desired peptide canbe attached, for example, to agarose, and the antibody-agarose complexis used in immunochromatography to purify peptides of the invention.See, Scopes, 1984, Protein Purification: Principles and Practice,Springer-Verlag New York, Inc., N.Y., Livingstone, 1974, Methods InEnzymology: Immunoaffinity Chromatography of Proteins 34:723-731.

5.3 Pharmaceutical Formulations and Methods of Treatment

The ApoA-I agonists of the invention can be used to treat any disorderin animals, especially mammals including humans, for which increasingserum HDL concentration, activating LCAT, and promoting cholesterolefflux and RCT is beneficial. Such conditions include, but are notlimited to hyperlipidemia, and especially hypercholesterolemia, andcardiovascular disease such as atherosclerosis (including treatment andprevention of atherosclerosis); restenosis (e.g., preventing or treatingatherosclerotic plaques which develop as a consequence of medicalprocedures such as balloon angioplasty); and other disorders, such asendotoxemia, which often results in septic shock.

The ApoA-I agonists can be used alone or in combination therapy withother drugs used to treat the foregoing conditions. Such therapiesinclude, but are not limited to simultaneous or sequentialadministration of the drugs involved.

For example, in the treatment of hypercholesterolemia oratherosclerosis, the ApoA-I agonist formulations can be administeredwith any one or more of the cholesterol lowering therapies currently inuse; e.g., bile-acid resins, niacin, and/or statins. Such a combinedregimen may produce particularly beneficial therapeutic effects sinceeach drug acts on a different target in cholesterol synthesis andtransport; i.e., bile-acid resins affect cholesterol recycling, thechylomicron and LDL population; niacin primarily affects the VLDL andLDL population; the statins inhibit cholesterol synthesis, decreasingthe LDL population (and perhaps increasing LDL receptor expression);whereas the ApoA-I agonists affect RCT, increase HDL, increase LCATactivity and promote cholesterol efflux.

In another embodiment, the ApoA-I agonists may be used in conjunctionwith fibrates to treat hyperlipidemia, hypercholesterolemia and/orcardiovascular disease such as atherosclerosis.

In yet another embodiment, the ApoA-I agonists of the invention can beused in combination with the anti-microbials and anti-inflammatoryagents currently used to treat septic shock induced by endotoxin.

The ApoA-I agonists of the invention can be formulated as peptides or aspeptide-lipid complexes which can be administered to subjects in avariety of ways to deliver the ApoA-I agonist to the circulation.Exemplary formulations and treatment regimens are described below.

5.3.1 ApoA-I Agonists and Peptide/Lipid Complex as the Active Ingredient

The ApoA-I agonist peptides can be synthesized or manufactured using anytechnique described in Section 5.2 and its subsections. Stablepreparations which have a long shelf life may be made by lyophilizingthe peptides--either to prepare bulk for reformulation, or to prepareindividual aliquots or dosage units which can be reconstituted byrehydration with sterile water or an appropriate sterile bufferedsolution prior to administration to a subject.

In certain embodiments, it may be preferred to formulate and administerthe ApoA-I agonist in a peptide-lipid complex. This approach has severaladvantages since the complex should have an increased half-life in thecirculation, particularly when the complex has a similar size anddensity to HDL, and especially the pre-β-1 or pre-β-2 HDL populations.The peptide-lipid complexes can conveniently be prepared by any of anumber of methods described below. Stable preparations having a longshelf life may be made by lyophilization--the co-lyophilizationprocedure described below being the preferred approach. The lyophilizedpeptide-lipid complexes can be used to prepare bulk for pharmaceuticalreformulation, or to prepare individual aliquots or dosage units whichcan be reconstituted by rehydration with sterile water or an appropriatebuffered solution prior to administration to a subject.

A variety of methods well known to those skilled in the art can be usedto prepare the peptide-lipid vesicles or complexes. To this end, anumber of available techniques for preparing liposomes orproteoliposomes may be used. For example, the peptide can be cosonicated(using a bath or probe sonicator) with appropriate lipids to formcomplexes. Alternatively the peptide can be combined with preformedlipid vesicles resulting in the spontaneous formation of peptide-lipidcomplexes. In yet another alternative, the peptide-lipid complexes canbe formed by a detergent dialysis method; e.g., a mixture of thepeptide, lipid and detergent is dialyzed to remove the detergent andreconstitute or form peptide-lipid complexes (e.g., see Jonas et al.,1986, Methods in Enzymol. 128:553-582).

While the foregoing approaches are feasible, each method presents itsown peculiar production problems in terms of cost, yield,reproducibility and safety. The applicants have developed a simplemethod for preparing peptide or protein-phospholipid complexes whichhave characteristics similar to HDL. This method can be used to preparethe ApoA-I peptide-lipid complexes, and has the following advantages:(1) Most or all of the included ingredients are used to form thedesigned complexes, thus avoiding waste of starting material which iscommon to the other methods. (2) Lyophilized compounds are formed whichare very stable during storage. The resulting complexes may bereconstituted immediately before use. (3) The resulting complexesusually need not be further purified after formation and before use. (4)Toxic compounds, including detergents such as cholate, are avoided.Moreover, the production method can be easily scaled up and is suitablefor GMP manufacture (i.e., in an endotoxin-free environment).

In accordance with the preferred method, the peptide and lipid arecombined in a solvent system which co-solubilizes each ingredient andcan be completely removed by lyophilization. To this end, solvent pairsmust be carefully selected to ensure co-solubility of both theamphipathic peptide and the lipid. In one embodiment, the protein(s) orpeptide(s) to be incorporated into the particles can be dissolved in anaqueous or organic solvent or mixture of solvents (solvent 1). The(phospho)lipid component is dissolved in an aqueous or organic solventor mixture of solvents (solvent 2) which is miscible with solvent 1, andthe two solutions are mixed. Alternatively, the peptide and lipid can beincorporated into a co-solvent system; i.e., a mixture of the misciblesolvents. A suitable proportion of peptide (protein) to lipids is firstdetermined empirically so that the resulting complexes possess theappropriate physical and chemical properties; i.e., usually (but notnecessarily) similar in size to HDL. The resulting mixture is frozen andlyophilized to dryness. Sometimes an additional solvent must be added tothe mixture to facilitate lyophilization. This lyophilized product canbe stored for long periods and will remain stable.

In the working examples describe infra, the peptide 210 (SEQ ID NO:210)and phospholipids were dissolved separately in methanol, combined, thenmixed with xylene before lyophilization. The peptide and lipid can bothbe added to a mixture of the two solvents. Alternatively, a solution ofthe peptide dissolved in methanol can be mixed with a solution of lipiddissolved in xylene. Care should be taken to eliminate salt from thesolvent system in order to avoid salting out the peptide. The resultingsolution containing the peptide and lipid cosolubilized inmethanol/xylene is lyophilized to form a powder.

The lyophilized product can be reconstituted in order to obtain asolution or suspension of the peptide-lipid complex. To this end, thelyophilized powder is rehydrated with an aqueous solution to a suitablevolume (often 5 mgs peptide/ml which is convenient for intravenousinjection). In a preferred embodiment the lyophilized powder isrehydrated with phosphate buffered saline or a physiological salinesolution. The mixture may have to be agitated or vortexed to facilitaterehydration, and in most cases, the reconstitution step should beconducted at a temperature equal to or greater than the phase transitiontemperature of the lipid component of the complexes. Within minutes, aclear preparation of reconstituted lipid-protein complexes results.

An aliquot of the resulting reconstituted preparation can becharacterized to confirm that the complexes in the preparation have thedesired size distribution; e.g., the size distribution of HDL. Gelfiltration chromatography can be used to this end. In the workingexamples described infra, a Pharmacia Superose 6 FPLC gel filtrationchromatography system was used. The buffer used contains 150 mM NaCl in50 mM phosphate buffer, pH 7.4. A typical sample volume is 20 to 200microliters of complexes containing 5 mgs peptide/ml. The column flowrate is 0.5 mls/min. A series of proteins of known molecular weight andStokes' diameter as well as human HDL are used as standards to calibratethe column. The proteins and lipoprotein complexes are monitored byabsorbance or scattering of light of wavelength 254 or 280 nm.

The ApoA-I agonists of the invention can be complexed with a variety oflipids, including saturated, unsaturated, natural and synthetic lipidsand/or phospholipids. Suitable lipids include, but are not limited to,small alkyl chain phospholipids, egg phosphatidylcholine, soybeanphosphatidylcholine, dipalmitoylphosphatidylcholine,dimyristoylphosphatidylcholine, distearoylphosphatidylcholine1-myristoyl-2-palmitoylphosphatidylcholine,1-palmitoyl-2-myristoylphosphatidylcholine,1-palmitoyl-2-stearoylphosphatidylcholine,1-stearoyl-2-palmitoylphosphatidylcholine, dioleoylphosphatidylcholinedioleophosphatidylethanolamine, dilauroylphosphatidylglycerolphosphatidylcholine, phosphatidylserine, phosphatidylethanolamine,phosphatidylinositol, sphingomyelin, sphingolipids,phosphatidylglycerol, diphosphatidylglycerol,dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol,distearoylphosphatidylglycerol, dioleoylphosphatidylglycerol,dimyristoylphosphatidic acid, dipalmitoylphosphatidic acid,dimyristoylphosphatidylethanolamine,dipalmitoylphosphatidylethanolamine, dimyristoylphosphatidylserine,dipalmitoylphosphatidylserine, brain phosphatidylserine, brainsphingomyelin, dipalmitoylsphingomyelin, distearoylsphingomyelin,phosphatidic acid, galactocerebroside, gangliosides, cerebrosides,dilaurylphosphatidylcholine, (1,3)-D-mannosyl-(1,3)diglyceride,aminophenylglycoside, 3-cholesteryl-6'-(glycosylthio)hexyl etherglycolipids, and cholesterol and its derivatives.

The Applicants have discovered that when the ApoA-I agonists of theinvention are complexed with sphingomyelin, all of the HDL of thepre-β-like particles is removed. Accordingly, in a preferred embodimentof the invention, the ApoA-I agonists are administered as a complex withsphingomyelin.

5.3.2 Methods of the Treatment

The ApoA-I peptide agonists or peptide-lipid complexes of the inventionmay be administered by any suitable route that ensures bioavailabilityin the circulation. This can best be achieved by parenteral routes ofadministration, including intravenous (IV), intramuscular (IM),intradermal, subcutaneous (SC) and intraperitoneal (IP) injections.However, other routes of administration may be used. For example,absorption through the gastrointestinal tract can be accomplished byoral routes of administration (including but not limited to ingestion,buccal and sublingual routes) provided appropriate formulations (e.g.,enteric coatings) are used to avoid or minimize degradation of theactive ingredient, e.g., in the harsh environments of the oral mucosa,stomach and/or small intestine. Alternatively, administration viamucosal tissue such as vaginal and rectal modes of administration may beutilized to avoid or minimize degradation in the gastrointestinal tract.In yet another alternative, the formulations of the invention can beadministered transcutaneously (e.g., transdermally), or by inhalation.It will be appreciated that the preferred route may vary with thecondition, age and compliance of the recipient.

The actual dose of ApoA-I agonists or peptide-lipid complex used willvary with the route of administration, and should be adjusted to achievecirculating plasma concentrations of 100 mg/l to 2 g/l. Data obtained inanimal model systems described herein show that the ApoA-I agonists ofthe invention associate with the HDL component, and have a projectedhalf-life in humans of about five days. Thus, in one embodiment, theApoA-I agonists can be administered by injection at a dose between 0.5mg/kg to 100 mg/kg once a week. In another embodiment, desirable serumlevels may be maintained by continuous infusion or by intermittentinfusion providing about 0.5 mg/kg/hr to 100 mg/kg/hr.

Toxicity and therapeutic efficacy of the various ApoA-I agonists can bedetermined using standard pharmaceutical procedures in cell culture orexperimental animals for determining the LD₅₀ (the dose lethal to 50% ofthe population) and the ED₅₀ (the dose therapeutically effective in 50%of the population). The dose ratio between toxic and therapeutic effectsis the therapeutic index and it can be expressed as the ratio LD₅₀/ED₅₀. ApoA-I peptide agonists which exhibit large therapeutic indicesare preferred.

5.3.3 Pharmaceutical Formulations

The pharmaceutical formulation of the invention contain the ApoA-Ipeptide agonist or the peptide-lipid complex as the active ingredient ina pharmaceutically acceptable carrier suitable for administration anddelivery in vivo. As the peptides may contain acidic and/or basictermini and/or side chains, the peptides can be included in theformulations in either the form of free acids or bases, or in the formof pharmaceutically acceptable salts.

Injectable preparations include sterile suspensions, solutions oremulsions of the active ingredient in aqueous or oily vehicles. Thecompositions may also contain formulating agents, such as suspending,stabilizing and/or dispersing agent. The formulations for injection maybe presented in unit dosage form, e.g., in ampules or in multidosecontainers, and may contain added preservatives.

Alternatively, the injectable formulation may be provided in powder formfor reconstitution with a suitable vehicle, including but not limited tosterile pyrogen free water, buffer, dextrose solution, etc., before use.To this end, the ApoA-I agonist may be lyophilized, or theco-lyophilized peptide-lipid complex may be prepared. The storedpreparations can be supplied in unit dosage forms and reconstitutedprior to use in vivo.

For prolonged delivery, the active ingredient can be formulated as adepot preparation, for administration by implantation; e.g.,subcutaneous, intradermal, or intramuscular injection. Thus, forexample, the active ingredient may be formulated with suitable polymericor hydrophobic materials (e.g., as an emulsion in an acceptable oil) orion exchange resins, or as sparingly soluble derivatives; e.g., as asparingly soluble salt form of the ApoA-I agonist.

Alternatively, transdermal delivery systems manufactured as an adhesivedisc or patch which slowly releases the active ingredient forpercutaneous absorption may be used. To this end, permeation enhancersmay be used to facilitate transdermal penetration of the activeingredient. A particular benefit may be achieved by incorporating theApoA-I agonists of the invention or the peptide-lipid complex into anitroglycerin patch for use in patients with ischemic heart disease andhypercholesterolemia.

For oral administration, the pharmaceutical compositions may take theform of, for example, tablets or capsules prepared by conventional meanswith pharmaceutically acceptable excipients such as binding agents(e.g., pregelatinised maize starch, polyvinylpyrrolidone orhydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystallinecellulose or calcium hydrogen phosphate); lubricants (e.g., magnesiumstearate, talc or silica); disintegrants (e.g., potato starch or sodiumstarch glycolate); or wetting agents (e.g., sodium lauryl sulfate). Thetablets may be coated by methods well known in the art. Liquidpreparations for oral administration may take the form of, for example,solutions, syrups or suspensions, or they may be presented as a dryproduct for constitution with water or other suitable vehicle beforeuse. Such liquid preparations may be prepared by conventional means withpharmaceutically acceptable additives such as suspending agents (e.g.,sorbitol syrup, cellulose derivatives or hydrogenated edible fats);emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles(e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetableoils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates orsorbic acid). The preparations may also contain buffer salts, flavoring,coloring and sweetening agents as appropriate. Preparations for oraladministration may be suitably formulated to give controlled release ofthe active compound.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in conventional manner. For rectal and vaginalroutes of administration, the active ingredient may be formulated assolutions (for retention enemas) suppositories or ointments.

For administration by inhalation, the active ingredient can beconveniently delivered in the form of an aerosol spray presentation frompressurized packs or a nebulizer, with the use of a suitable propellant,e.g., dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol the dosage unit may be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof e.g. gelatin for use in an inhaler or insufflator may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

The compositions may, if desired, be presented in a pack or dispenserdevice which may contain one or more unit dosage forms containing theactive ingredient. The pack may for example comprise metal or plasticfoil, such as a blister pack. The pack or dispenser device may beaccompanied by instructions for administration.

5.4 Other Uses

The ApoA-I agonists of the invention can be used in assays in vitro tomeasure serum HDL, e.g., for diagnostic purposes. Because the ApoA-Iagonists associate with the HDL component of serum, the agonists can beused as "markers" for the HDL population. Moreover, the agonists can beused as markers for the subpopulation of HDL that are effective in RCT.To this end, the agonist can be added to or mixed with a patient serumsample; after an appropriate incubation time, the HDL component can beassayed by detecting the incorporated ApoA-I agonist. This can beaccomplished using labeled agonists (eq., radiolabels, fluorescentlabels, enzyme labels, dyes, etc.), or by immunoassays using antibodies(or antibody fragments) specific for the agonist.

Alternatively, labeled agonist can be used in imaging procedures (e.g.,CAT scans, MRI scans) to visualize the circulatory system, or to monitorRCT, or to visualize accumulation of HDL at fatty streaks,atherosclerotic lesions, etc. (where the HDL should be active incholesterol efflux).

6. EXAMPLE

Synthesis of Peptide Agonists of ApoA-I

The peptides described in TABLE IX (Section 8.3, infra) were synthesizedand characterized as described in the subsections below. The peptideswere also analyzed structurally and functionally as described inSections 7 and 8, infra.

6.1 Synthesis of Core Peptides

Peptides were synthesized on solid phase according to the Merrifieldtechnique (Merrifield, 1969, J. Am. Chem. Soc. 85:2149-2154) using 0.25mmol ρ-alkoxybenzylalcohol resin (HMP resin) (Wang, 1973, J. Am. Chem.Soc. 95:1328-1333) and Fmoc chemistry. All syntheses were carried out onan Applied Biosystems ABI model 430A automated peptide synthesizer(Perkin-Elmer, Foster City, Calif.). The solvation and activation timesused for each coupling cycle are shown in TABLE V below:

                                      TABLE V                                     __________________________________________________________________________    SINGLE COUPLE ACTIVATOR CYCLES                                                CYCLE                                                                             DESIGNATED                                                                            DISSOLVING  ACTIVATION                                                                           TRANSFER                                       NAME                                                                              AMINO ACIDS                                                                           SOLVENT                                                                              TIME TIME   TIMES*                                         __________________________________________________________________________    afmc 31                                                                           Asn (trt),                                                                            ˜0.4 ml DCM                                                                    ˜7 min.                                                                      ˜51 min.                                                                       1 = 50 sec.                                        His (trt),                                                                            ˜1.2 ml NMP  2 = 36 sec.                                        Lys (Boc), Trp                                                                        ˜1.0 ml                                                                 HOBt/NMP                                                          afmc 32                                                                           Arg (Pmc),                                                                            ˜0.8 ml DCM                                                                    ˜32 min.                                                                     ˜51 min.                                                                       1 = 60 sec.                                        Gln (trt), Aib                                                                        ˜1.2 ml NMP  2 = 40 sec.                                                ˜1.0 ml                                                                 HOBt/NMP                                                          afmc 33                                                                           Ala, Asp (OtBu),                                                                      ˜0.4 ml DCM                                                                    ˜4 min.                                                                      ˜36.5                                                                          1 = 38 sec.                                        Glu (OtBu), Gly,                                                                      ˜0.8 ml NMP                                                                         min.   2 = 27 sec.                                        Ile, Leu,                                                                             ˜0.1 ml                                                         Met, Phe, Pro                                                                         HOBt/NMP                                                          afmc 34                                                                           Val     ˜0.4 ml DCM                                                                    ˜4 min.                                                                      ˜61.5                                                                          1 = 38 sec.                                                ˜0.8 ml NMP                                                                         min.   2 = 27 sec.                                                ˜0.1 ml                                                                 HOBt/NMP                                                          __________________________________________________________________________     *1 = Transfer from Cartridge to Activator.                                    2 = Transfer from Activator to Cartridge.                                     DCC is dicyclohexylcarbodiimide                                               BOC is tbutyloxycarbonyl                                                      HOBt is 1hydroxybenzotriazole                                                 Pmc is pentamethylchroman6-sulfonyl                                           NMP is Nmethylpyrrolidone                                                     OtBu is tbutyl ester                                                          trt is trityl                                                            

The resins were washed with NMP between each coupling step. The protocolfor one synthesis cycle is shown below in TABLE VI:

                  TABLE VI                                                        ______________________________________                                        COUPLING PROTOCOL FOR ONE SYNTHESIS CYCLE                                            OPERATION      TIME (min.)                                             ______________________________________                                        1.       Deprotection (10% piperdine                                                                    20                                                           in NMP)                                                              2.       Wash (NMP)        5                                                  3.       Couple (4 equiv. Fmoc-amino                                                                    61                                                           acid-HOBT ester in NMP,                                                       preactivated 50 min.)                                                4.       Wash              3                                                  5.       Resin Sample (optional)                                                                         3                                                           TOTAL            92                                                  ______________________________________                                    

All amino acids except Fmoc-β-(1-naphthyl)alanine were coupled in thismanner. Fmoc-β-(1-naphthyl)alanine was coupled manually. For manualcoupling, 1 mmol Fmoc-β-(1-naphthyl)alanine and 1 mmol2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate(TBTU) were dissolved in 5 ml NMP and mixed with the peptide-resin.

Thereafter, 2 mmol of N-ethyldiisopropylamine were added, the mixtureshaken for 2 hours and the peptide-resin washed 6 times with 10 ml NMP.The coupling efficiency was monitored using the Kaiser Test (Kaiser,1970, Anal. Biochem. 34:59577), and the coupling repeated if necessary.After coupling of naphthylalanine, the remainder of the synthesis wasperformed automatically as described above.

6.2 Synthesis of Peptide Amides

Where indicated in TABLE IX (Section 8.3, infra), peptide amides weresynthesized using a Rink amide resin containing the Fmoc-Rink amidehandle 4-(2',4'-dimethylphenyl)-Fmoc-phenoxymethyl (Rink, 1987,Tetrahedron Lett. 28:3787-3790) and the synthesis protocols described inSection 6.1, supra.

6.3 Synthesis of N-Terminal Acylated Peptides

Where indicated in TABLE IX (Section 8.3, infra), N-terminal acylatedforms of the peptides were prepared by exposing the resin-bound peptideprepared as described in Section 6.1 or 6.2, supra, to an appropriateacylating agent.

For N-terminal acetylated peptides, 15 ml of acetic anhydride solution(10% v/v in NMP) was added to each 1 g of resin-bound peptide, themixture shaken for 5 min. and the resin recovered by filtration. Therecovered resin was washed three times with NMP (15 ml) and three timeswith ethanol (15 ml).

6.4 CLeavage and Deprotection

Following synthesis, the peptides described in Sections 6.1, 6.2 and6.3, supra, were cleaved from the resin and deprotected with a cleavagesolution containing 92.5% trifluroacetic acid (TFA)/3.75% anisole/3.75%dodecanthiol (v/v/v). To effect cleavage, 10 ml of cleavage solution wasadded to 0.25 mmol peptide resin and stirred for 1.5 hours at roomtemperature. The resin was removed via filtration and thecleaved/deprotected peptide precipitated with diethyl ether, washed withether and dried in vacuo.

The cleavage cocktail for peptides containing Trp (W), as well as forpeptide amides, was composed of 86.5% TFA, 4.5% H₂ O, 4.5%1,2-ethanedithiol, 4.5% anisole and 3% phenol.

6.5 Purification

The crude, cleaved peptides of Section 6.4 were purified by reversephase HPLC. The purity of each peptide was confirmed by differentanalytical techniques (analytical HPLC, capillary electrophoresis).Capillary electrophoreses were carried out on fused silica capillariesof 70 cm length and an internal diameter of 75 μm (Thermo SeparationProducts). The separations were performed at 25° C., 15 kV, run time 35min., in two different buffer systems: Buffer 1 (20 mM Na₂ B₄ O₇, pH9.2) and Buffer 2 (10 mM Na₂ HPO₄, pH 2.5). HPLC separations werecarried out on Nucleosil 7C18 or Nucleosil 7C4 columns (Macherey andNagel, Germany), 250×21 mm, at a flow rate of 8 ml/min. The gradientelution was performed using a mixture of 0.1% TFA in water (Solvent A)and 0.1% TFA in acetonitrile (Solvent B). The gradients used wereadjusted to meet the needs of each peptide.

6.6 Characterization

The mass and amino acid analysis of the purified peptides described inSection 6.5 were confirmed via mass spectrometry and amino acidanalysis, respectively, as described below. Edman degradation was usedfor sequencing.

6.6.1 LC-MS

A standard commercially available triple stage quadruple massspectrometer (model TSQ 700; Finnigan MAT, San Jose Calif., U.S.A.) wasused for mass determination. A pneumatically assisted electrospray (ESI)interface was used for sample introduction to the atmospheric pressureionization source of the mass spectrometer. The interface sprayer wasoperated at a positive potential of 4.5 kV. The temperature of the steelcapillary was held at 200° C. whereas the manifold was at 70° C.Positive ions generated by this ion evaporation process entered theanalyzer of the mass spectrometer. The multiplier was adjusted to 1000V. The analyzer compartment of the mass spectrometer was at 4E-6. Allacquisitions were performed at resolution <1u.

Peptides were analyzed by direct infusion of the purified peptides usingan ABI (Applied Biosystems) microbore system consisting of a syringepump (model 140B), an UV detector (model 785A) and an oven/injector(model 112A). The solvent system consisted of water (solvent A) andacetonitrile (solvent B), each containing 0.1% TFA. Peptides wereinfused using either a gradient or isocratic conditions and eluted froman Aquapore C18 column. The flow rate was typically 300 μl/min.Concentration of each peptide was about 0.03 mg/ml, 20 μl of which wasinjected (e.g., 30 pmol).

Full scan MS experiments were obtained by scanning quadruple 1 from m/z500-1500 in 4 s. Data were acquired using an Alpha DEC station and wereprocessed using the software package provided by Finnigan MAT(BIOWORKS).

6.6.2 Amino Acid Analysis

Amino acid analysis was performed on an ABI (Applied Biosystems) 420Amino Acid Analyzer. This system consists of three modules: a hydrolysisand derivatisation instrument, a reverse-phase HPLC and a data system.Peptide sample were applied (3 times in triplicate) on porous glassslides and subsequently hydrolyzed under gas phase conditions (155° C.,90 min.). After removal of the HCL, the resulting amino acids wereconverted to PTC-AA (Phenylthiocarbamoyl-amino acids) using PITC(Phenylisothiocyanate). After transfer to the HPC sample loop theresulting mixtures were fractionated on an Aquapore C18 column using thegradient mode (Solvent A: 50 mmol ammonium acetate (NH₄ Ac), pH 5.4, inwater; Solvent B: 32 mmol of sodium acetate (NaOAc) in aqueousacetonitrile) under conditions of temperature control. The HPLC datawere processed by the software package provided by Applied Biosystems.Quantification was performed relative to a peptide standard delivered byApplied Biosystems.

6.7 Synthesis of Branched Networks

Tetrameric-core peptidyl resin and trimeric-core peptidyl resin aresynthesized as described in Demoor et al., 1996, Eur. J. Biochem.239:74-84. The tetrameric and trimeric core matrix still linked to the4-methyl benzhydrylamine resin is then used as initial peptidyl-resinfor automated synthesis of core peptides as previously described.

Branched networks containing helical segments of different amino acidcompositions can be synthesized using orthogonal synthesis andprotecting strategies well known in the art.

7. EXAMPLE

Structural and Lipid Binding Analysis of ApoA-I Peptides

The structural and lipid binding characteristics of the purifiedpeptides synthesized as described in Section 6, supra, were determinedby circular dichroism (CD), fluorescence spectroscopy and nuclearmagnetic resonance (NMR).

7.1 Circular Dichroism

This Example describes a preferred method for determining the degree ofhelicity of the core peptides of the invention both free in buffer andin the presence of lipids.

7.1.1 Experimental Method

Far UV circular dichroism spectra were recorded between 190 and 260 nm(in 0.5 nm or 0.2 nm increments) with a AVIV62DS spectrometer (AVIVAssociates, Lakewood, N.J., U.S.A.) equipped with a thermoelectric cellholder and sample changer. The instrument was calibrated with(+)-10-camphoric acid. Between one and three scans were collected foreach sample, using 10 cm, 5 cm, 1 cm and 0.1 cm path length quartzSuprasil cells, respectively, for peptide concentrations of 10⁻⁷ M to10⁻⁴ M. The bandwidth was fixed at 1.5 nm and the scan speed to 1 s perwavelength step. The reported data are the mean of at least 2 or 3independent measurements.

After background substraction, spectra were converted to molarellipticity (θ) per residue in deg. cm⁻² dmol⁻¹. The peptideconcentration was determined by amino acid analysis and also byabsorption spectrometry on a Perkin Elmer Lambda 17 UV/Visiblespectrophotometer when the peptide contained a chromophore (tryptophane,dansyl, naphtylalanine).

CD spectra were obtained with free, unbound peptide (5 μM in 5 mMphosphate buffer, pH 7.4); with peptide-SUV complexes (20:1 EPC:Chol.,Ri=50); with peptide-micelle complexes(1-myristoyl-2-hydroxy-sn-glycero-3-phosphatidyl choline, Ri=100); andwith free, unbound peptide in the presence of 2,2,2-trifluoroethanol(TFE) (5 μM peptide, 90% vol TFE).

The SUVs were obtained by dispersing the lipids (10 mM, 20:1 EPC:Chol.,Avanti Polar Lipids, AL) in phosphate buffer (5 mM, pH 7.4) withbubbling N₂ for 5 min., followed by sonication (1.5 hr.) in a bathsonicator. The homogeneity of the preparation was checked by FPLC.

The micelles were obtained by dispersing the lipid (6 mM1-myristoyl-2-hydroxy-sn-glycero-3-phosphatidyl choline, Avanti PolarLipids, AL) in phosphate buffer (5 mM, pH 7.4) with bubbling N₂ for 5min., followed by vortexing.

To obtain the peptide-SUV complexes, SUVs were added to the peptide (5μM in 5 mM phosphate buffer, pH 7.4) at a phospholipid-peptide molarratio (Ri) of 100.

To obtain the peptide-micelle complexes, micelles were added to thepeptide (5 μM in 5 mM phosphate buffer, pH 7.4) at a Ri of 100.

All spectra were recorded at 37° C. The stability of peptide 210 (SEQ IDNO:210) as a function of temperature (both free in buffer and inmicelles) was determined by recording spectra at a series of differenttemperatures.

The degree of helicity of peptide 210 (SEQ ID NO:210) as a function ofconcentration was also determined.

7.1.2 Helicity Determination

The degree of helicity of the peptides in the various conditions wasdetermined from the mean residue ellipticity at 222 nm (Chen et al.,1974, Biochemistry 13:3350-3359) or by comparing the CD spectra obtainedto reference spectra available on databases (16 helical referencespectra from Provencher & Glockner, 1981, Biochemistry 20:33-37;denatured protein reference spectra from Venyaminov et al., 1993, Anal.Biochem. 214:17-24) using the CONTIN curve-fitting algorithm version2DP, CD-1 pack (August 1982) (Provencher, 1982, Comput. Phys. Commun.27:213-227, 229-242). Acceptable fit was determined using thestatistical analysis methodology provided by the CONTIN algorithm. Theerror of all methods was ±5% helicity.

7.1.3 Results

The degree of helicity (%) of the free, unbound peptides (free), thepeptide-SUV complexes (SUVs), the peptide-micelle complexes (mics) andthe peptide-TFE solution (TFE) are reported in TABLE IX, Section 8.3,infra.

Peptide 210 (SEQ ID NO:210) contains significant α-helical structure(63% helicity) in micelles. Moreover, the α-helical structure iscompletely stable over a temperature range of 5°-45° C. (data notshown). The helicity of peptide 210 (SEQ ID NO:210) also increases inthe presence of TFE, which is a solvent that, due to having asignificantly lower dielectric constant (ε=26.7) than water (ε=78.4),stabilizes α-helices and intrapeptide hydrogen bonds at concentrationsbetween 5-90% (v/v).

Referring to TABLE IX, Section 8.3, infra, it can be seen that thosepeptides which exhibit a high degree of LCAT activation (≧38%) generallypossess significant α-helical structure in the presence of lipids (≧60%helical structure in the case of unblocked peptides containing 22 ormore amino acids or blocked peptides containing 18 or fewer amino acids;≧40% helical structure in the case of unblocked peptides containing 18or fewer amino acids), whereas peptides which exhibit little or no LCATactivation possess little α-helical structure. However, in someinstances, peptides which contain significant α-helical structure in thepresence of lipids do not exhibit significant LCAT activation. As aconsequence, the ability of the core peptides of the invention to adoptan α-helical structure in the presence of lipids is considered acritical feature of the core peptides of the invention, as the abilityto form an α-helix in the presence of lipids appears to be aprerequisite for LCAT activation.

7.2 Fluorescence Spectroscopy

The lipid binding properties of the peptides synthesized in Section 6,supra, were tested by fluorescence measurements with labeled peptides,in the present case Tryptophane (Trp or W) or Naphtylalanine (Nal). Thefluorescence spectra were recorded on a Fluoromax from Spex (Jobin-Yvon)equipped with a Xenon lamp of 150 W, two monochromators (excitation andemission), a photomultiplier R-928 for detection sensitive in the red upto 850 nm and a thermoelectric magnetic stirred cell holder. QuartzSuprasil cuvettes were used for measurements in the micromolarconcentration range. A device of variable slits (from 0.4 to 5 nm)allows modulation of the incident and emitted intensities according tothe concentration of peptide used. The reported values are in generalthe average of between 2 to 4 spectra. The peptide concentration isdetermined by absorption spectrometry on a Philips PU 8800 using theabsorption band of the Trp (ε₂₈₀ nm =5,550 M⁻¹ cm⁻¹ in Tris buffer) orthe Nal (ε₂₂₄ nm =92,770 M⁻¹ cm⁻¹ in methanol).

Fluorescence spectra of the peptides were recorded between 290 nm and450 nm in Tris-HCl buffer (20 mM, pH=7.5), in the presence and absenceof lipidic vesicles. The small unilamellar vesicles were formed afterrehydration in buffer of the lyophilized phospholipids, dispersion andtip sonification under a N₂ stream. The lipids used were either EggPC/Chol. (20:1) or POPC/Chol. (20:1). The spectra were recorded at apeptide concentration of 2 μM and at a temperature of 37° C. Thefluorescence reference standard in the case of Trp wasN-acetyltryptophanylamide (NATA).

Lipid binding studies were done through progressive lipidic vesicleaddition to the peptide in solution at 2 μM (slits:5 nm in excitationand 1.5 nm in emission). Dilution effects were taken into account forthe fluorescence intensity determination. The lipid concentrations werevaried from 10 to 600 μM and the molar ratio of lipid to peptide (Ri)was varied from 5 to 300. The wavelength of excitation was set at 280 nmfor both Trp and Nal.

7.2.1 Fluorescence Spectral Analysis

The data were directly recorded and treated by an IBM-PC linked to thespectrofluorimeter through the DM3000F software from Spex. The spectrawere corrected by substraction of the solvent contribution and byapplication of a coefficient given by the constructor taking intoaccount the variation of the photomultiplier response versus thewavelength.

The fluorescence spectra of the peptides were characterized by thewavelength at their maximum of fluorescence emission and by theirquantum yield compared to NATA in the case of peptides labeled with atryptophane. The process of binding to lipids was analyzed bycalculating the shift of the wavelength at the maximum of fluorescenceemission, (λ_(max)), and the variation of the relative fluorescenceintensity of emission versus the lipid concentration. The relativefluorescence intensity is defined as the following ratio:(I-I₀).sub.λmax /I₀λmax. I and I₀ are both measured at the (λ_(max))corresponding to the initial free state of the peptide, i.e., withoutlipids. I is the intensity at a defined lipid to peptide ratio and I₀ isthe same parameter measured in absence of lipids. The absence of thesevariations is relevant of the absence of interactions of the peptideswith the lipids.

7.2.2 Results and Discussion

The lipid binding properties of peptide 199 (SEQ ID NO:199), which issimilar in primary sequence to peptide 210 (SEQ ID NO:210) except thatit contains a W (Trp) residue at position 10, are presented in TABLEVII.

                  TABLE VII                                                       ______________________________________                                        BINDING PROPERTIES OF PEPTIDE 199 (SEQ ID NO:199)                             TO LIPIDIC VESICLES AS MEASURED BY FLUORESCENCE                               Lipid:Peptide                                                                 Molar Ratio (Ri) I/I.sub.o                                                                           .sub.max (nm)                                          ______________________________________                                         0                0    348                                                     5                8    344                                                    10                8    339                                                    30               18    328                                                    60               22                                                           100              27    326                                                    200              41    325                                                    ______________________________________                                    

In buffer at a concentration of 2 μm, the maximum of the tryptophanefluorescence emission (λ_(max)) of peptide 199 (SEQ ID NO:199) is 348nm. This corresponds to a tryptophane which is relatively exposed to theaqueous environment when compared to NATA (λ max=350 nm). Peptide 199(SEQ ID NO:199) binds very effectively to EPC/Chol (20:1) smallunilamellar vesicles as demonstrated by the burying of the tryptophane(the wavelength for the tryptophane maximum fluorescence emission shiftsfrom 348 nm to 325 nm) and the high fluorescence intensity exaltation(see Table VII). The burying of the tryptophane residue is maximal for alipid to peptide molar ratio of about 100.

Other peptides which exhibited a high degree of helicity in the presenceof lipids (≧60% for unblocked peptides of ≧22 amino acids, or blockedpeptides of ≦18 amino acids; ≧40% for unblocked peptides of ≦18 aminoacids) as measured by circular dichroism as disclosed in Section 7.1,supra, also demonstrated good lipid binding. Of course, among all thepeptides selected by the circular dichroism screening, only the onesthat could be followed by fluorescence were tested for their lipidbinding properties.

7.3 Nuclear Magnetic Resonance (NMR)

This Example describes an NMR method for analyzing the structure of thecore peptides of the invention.

7.3.1 NMR Sample Preparation

Samples were prepared by dissolving 5 mg of peptide in 90% H₂ O/10% D₂ Ocontaining trace amounts of 2,2-Dimethyl-2-sila-5-pentane sulfonate(DSS) as an internal chemical shift reference. Some of the samplescontained trifluoroethanol (TFE) (expressed as % vol). The total samplevolume was 500 μl and the concentration of peptide was approximately 5mM.

7.3.2 NMR Spectroscopy

¹ H NMR spectra were acquired at 500 MHz using a Bruker DRX500spectrometer equipped with a B-VT2000 temperature control unit. One andtwo-dimensional experiments were recorded using standard pulsesequences. (Two Dimensional NMR Spectroscopy, Eds. W. R. Croasmun and RM K Carlson, 1994, VCH Publishers, New York, U.S.A.). Water suppressionwas achieved with low power presaturation for 2 sec. Two-dimensionalexperiments were carried out in the phase sensitive mode using timeproportional phase incrementation (TPPI) and a spectral width of 6000 Hzin both dimensions. Typically, 40 scans were co-added for 400 t₁increments with 2048 data points. Data were processed using FELIX95software (Molecular Simulations) on an INDIGO2 workstation (SiliconGraphics). Data were zero-filled to give a 2K×2K data matrix andapodized by a 45° shifted squared sine-bell function.

7.3.3 NMR Assignment

Complete proton resonance assignments were obtained by applying thesequential assignment technique using DQFCOSY, TOCSY and NOESY spectraas described in the literature (Wuthrich, NMR of Proteins and NucleicAcids, 1986, John Wiley & Sons, New York, U.S.A.). Secondary chemicalshifts were calculated for HN and Hα protons by subtracting thetabulated random coil chemical shifts (Wishart and Sykes, 1994, Method.Enz. 239:363-392) from the corresponding experimental values.

7.3.4 Results and Discussion

General Consideration

Amphipathic helical peptides tend to aggregate in aqueous solutions atthe high concentrations necessary for NMR spectroscopy, making itdifficult to obtain high resolution spectra. TFE is known to solubilizepeptides, and in addition stabilizes helical conformations of peptideshaving helical propensity. The findings from NMR spectroscopy aredemonstrated for peptide 210 (SEQ ID NO:210) as a representativeexample. The consensus 22-mer of Segrest (SEQ ID NO:75) was studied incomparison.

Secondary Chemical Shifts

Proton chemical shifts of amino acids depend both on the type of residueand on the local secondary structure within a peptide or protein(Szlagyi, 1995, Progress in Nuclear Magnetic Resonance Spectroscopy27:325-443). Therefore, identification of regular secondary structure ispossible by comparing experimental shifts with tabulated values forrandom coil conformation.

Formation of an α-helix typically results in an up-field (negative)shift for the Hα resonance. Observation of an upfield Hα shift forseveral sequential residues is generally taken as evidence of a helicalstructure. The Hα secondary shifts for peptide 210 (SEQ ID NO:210) in25% TFE at 295K show a significant negative shift for residues 4 through15 (FIG. 7A), demonstrating a highly helical conformation. Smalldifferences are observed in the Hα chemical shifts of the consensus22-mer (SEQ ID NO:75) compared to peptide 210 (SEQ ID NO:210).

The chemical shifts of amide hydrogens of amino acid residues residingin regions of α-helix are also shifted upfield with respect to thechemical shifts observed for random coil. In addition, a periodicity ofthe HN shifts can be observed, and it reflects the period of the helicalturns. The amplitude of the shift variation along the sequence isrelated to the amphipathicity of a helical peptide. A higher hydrophobicmoment leads to a more pronounced oscillation (Zhou et al., 1992, J. Am.Chem. Soc. 114:4320-4326). The HN secondary shifts for peptide 210 (SEQID NO:210) in 25% TFE at 295K show an oscillatory behavior in agreementwith the amphipathic nature of the helix (FIG. 7B).

The amino acid replacements lead to a more pronounced periodicity alongthe entire sequence (FIG. 7B). The pattern clearly reflects the strongeramphipathic nature of peptide 210 (SEQ ID NO:210) as compared toSegrest's consensus 22-mer (SEQ ID NO:75). The existence of 4-5 helicalturns can be discerned.

The secondary shift of an amide proton is influenced by the length ofthe hydrogen bond to the carbonyl oxygen one turn away from the helix.Therefore, the periodicity of observed chemical shift values reflectsdifferent hydrogen bond lengths. This difference is associated with anoverall curved helical shape of the helix backbone. The hydrophobicresidues are situated on the concave side. The secondary shifts ofpeptide 210 (SEQ ID NO:210) indicate a curved α-helical conformation.

8. EXAMPLE

LCAT Activation Assay

The peptides synthesized as described in Section 6, supra, were analyzedin vitro for their ability to activate LCAT. In the LCAT assay,substrate vesicles (small unilamellar vesicles or "SUVs") composed ofegg phophatidylcholine (EPC) or 1-Palmitoyl-2-oleyl-phosphatidylcholine(POPC) and radiolabelled cholesterol are preincubated with equivalentmasses either of peptide or ApoA-I (isolated from human plasma). Thereaction is initiated by addition of LCAT (purified from human plasma).Native ApoA-I, which was used as positive control, represents 100%activation activity. "Specific activity" (i.e., units of activity (LCATactivation)/unit of mass) of the peptides can be calculated as theconcentration of peptide that achieves maximum LCAT activation. Forexample, a series of concentrations of the peptide (e.g., a limitingdilution) can be assayed to determine the "specific activity" for thepeptide--the concentration which achieves maximal LCAT activation (i.e.,percentage conversion of cholesterol to cholesterol ester) at a specifictimepoint in the assay (e.g., 1 hr.). When plotting percentageconversion of cholesterol at, e.g., 1 hr., against the concentration ofpeptide used, the "specific activity" can be identified as theconcentration of peptide that achieves a plateau on the plotted curve.

8.1 Preparation of Substrate Vesicles

The vesicles used in the LCAT assay are SUVs composed of eggphosphatidylcholine (EPC) or 1-palmitoyl-2-oleyl-phosphatidylcholine(POPC) and cholesterol with a molar ratio of 20:1. To prepare a vesiclestock solution sufficient for 40 assays, 7.7 mg EPC (or 7.6 mg POPC; 10μmol), 78 μg (0.2 μmol) 4-¹⁴ C-cholesterol, 116 μg cholesterol (0.3μmol) are dissolved in 5 ml xylene and lyophilized. Thereafter 4 ml ofassay buffer is added to the dry powder and sonicated under nitrogenatmosphere at 4° C. Sonication conditions: Branson 250 sonicator, 10 mmtip, 6×5 minutes; Assay buffer: 10 mM Tris, 0.14 M NaCl, 1 mM EDTA, pH7.4). The sonicated mixture is centrifuged 6 times for 5 minutes eachtime at 14,000 rpm (16,000× g) to remove titanium particles. Theresulting clear solution is used for the enzyme assay.

8.2 Purification of LCAT

For the LCAT purification, dextran sulfate/Mg²⁺ treatment of humanplasma is used to obtain lipoprotein deficient serum (LPDS), which issequentially chromatographed on Phenylsepharose, Affigelblue,ConcanavalinA sepharose and anti-ApoA-I affinity chromatography, assummarized for a representative purification in TABLE VIII, below:

                                      TABLE VIII                                  __________________________________________________________________________    LCAT PURIFICATION                                                                  Total Volume                                                                         Total Protein                                                                       Total Activity                                                                        Yield                                                                            Purification                                     Fraction                                                                           (ml)   (mg)  (nmol CE/mg*hr)                                                                       (%)                                                                              (fold)                                           __________________________________________________________________________    Plasma                                                                             550    44550 63706                                                       LPDS 500    31000 62620   98 1.4                                              Phenyl                                                                             210    363   51909   82 100                                              sepharose                                                                     Affigel                                                                             95    153   25092   39 115                                              blue                                                                          ConA  43    36    11245   18 220                                              sepharose                                                                     Anti-A-I                                                                           120    3.5    5500    9 1109                                             Affinity                                                                      __________________________________________________________________________

8.2.1 Preparation of LPDS

To prepare LPDS, 500 ml plasma is added to 50 ml dextran sulfate(MW=500000) solution. Stir 20 minutes. Centrifuge for 30 minutes at 3000rpm (16,000×g) at 4° C. Use supernatant (LPDS) for further purification(ca. 500 ml).

8.2.2 Phenylsepharose Chromatography

The following materials and conditions were used for the phenylsepharosechromatography.

solid phase: Phenylsepharose fast flow, high subst. grade, Pharmacia

column: XK26/40, gel bed height: 33 cm, V=ca. 175 ml

flow rates: 200 ml/hr (sample)

wash: 200 ml/hr (buffer)

elution: 80 ml/hr (distilled water)

buffer: 10 mM Tris, 140 mM NaCl, 1 mM EDTA pH7.4, 0.01% sodium azide.

Equilibrate the column in Tris-buffer, add 29 g NaCl to 500 ml LPDS andapply to the column. Wash with several volumes of Tris buffer until theabsorption at 280 nm wavelength is approximately at the baseline, thenstart the elution with distilled water. The fractions containing proteinare pooled (pool size: 180 ml) and used for Affigelblue chromatography.

8.2.3 Affigelblue Chromatography

The Phenylsepharose pool is dialyzed overnight at 4° C. against 20 mMTris-HCl, pH 7.4, 0.01% sodium azide. The pool volume is reduced byultrafiltration (Amicon YM30) to 50-60 ml and loaded on an Affigelbluecolumn.

solid phase: Affigelblue, Biorad, 153-7301 column, XK26/20, gel bedheight: ca. 13 cm; column volume: approx. 70 ml.

flow rates: loading: 15 ml/h wash: 50 ml/h

Equilibrate column in Tris-buffer. Apply Phenylsepharose pool to column.Start in parallel to collect fractions. Wash with Tris-buffer. Thepooled fractions (170 ml) were used for ConA chromatography.

8.2.4 ConA Chromatography

The Affigelblue pool was reduced via Amicon (YM30) to 30-40 ml anddialyzed against ConA starting buffer (1 mM Tris HCl pH7.4; 1 mM MgCl₂,1 mM MnCl₂, 1 mM CaCl₂, 0.01% Na-azide) overnight at 4° C.

solid phase: ConA sepharose (Pharmacia)

column: XK26/20, gel bed height: 14 cm (75 ml)

flow rates: loading 40 ml/h washing (with starting buffer): 90 ml/helution: 50 ml/h, 0.2M Methyl-α-D-mannoside in 1 mM Tris, pH 7.4.

The protein fractions of the mannoside elutions were collected (110 ml),and the volume was reduced by ultrafiltration (YM30) to 44 ml. The ConApool was divided in 2 ml aliquots, which are stored at -20° C.

8.2.5 Anti-ApoA-I Affinity Chromatography

Anti-ApoA-I affinity chromatography was performed on Affigel-Hz material(Biorad), to which the anti-ApoA-I abs have been coupled covalently.

column: XK16/20, V=16 ml. The column was equilibrated with PBS pH 7.4.Two ml of the ConA pool was dialyzed for 2 hours against PBS beforeloading onto the column.

flow rates: loading: 15 ml/hour washing (PBS) 40 ml/hour.

The pooled protein fractions (V=14 ml) are used for LCAT assays.

The column is regenerated with 0.1 M. Citrate buffer (pH 4.5) to elutebound A-I (100 ml), and immediately after this procedure reequilibratedwith PBS.

8.3 Results

The results of the LCAT activation assay are presented in TABLE IX,infra.

                                      TABLE IX                                    __________________________________________________________________________    LCAT ACTIVATION EXHIBITED BY EXEMPLARY CORE PEPTIDES                                                        ACTIVITY                                                                            He (%)                                                                            He (%)                                                                            He (%)                                                                            He (%)                        PEPTIDE       AMINO ACID SEQUENCE                                                                           (%) LCAT                                                                            free                                                                              mica                                                                              SUVs                                                                              TFE                           __________________________________________________________________________     1   (SEQ ID NO: 1)                                                                         PVLDLFRELLNELLEZLKQKLK                                                                        120%  77  85  81  69                             2   (SEQ ID NO: 2)                                                                         GVLDLFRELLNELLEALKQKLKK                                                                       105%                                             3   (SEQ ID NO: 3)                                                                         PVLDLFRELLNELLEWLKQKLK                                                                        98%   70  95  80  95                             4   (SEQ ID NO: 4)                                                                         PVLDLFRELLNELLEALKQKLK                                                                        93%   80  95  97  94                             5   (SEQ ID NO: 5)                                                                         pVLDLFRELLNELLEALKQKLKK                                                                       90%                                              6   (SEQ ID NO: 6)                                                                         PVLDLFRELLNEXLEALKQKLK                                                                        80%   57  93  70  99                             7   (SEQ ID NO: 7)                                                                         PVLDLFKELLNELLEALKQKLK                                                                        83%   77  89  85  73                             8   (SEQ ID NO: 8)                                                                         PVLDLFRELLNEGLEALKQKLK                                                                        83%   20  90  61  93                             9   (SEQ ID NO: 9)                                                                         PVLDLFRELGNELLEALKQKLK                                                                        83%                                              10  (SEQ ID NO: 10)                                                                        PVLDLFRELLNELLEAZKQKLK                                                                        79%   60  87  70  71                             11  (SEQ ID NO: 11)                                                                        PVLDLFKELLQELLEALKQKLK                                                                        72%                                              12  (SEQ ID NO: 12)                                                                        PVLDLFRELLNELLEAGKQKLK                                                                        70%                                              13  (SEQ ID NO: 13)                                                                        GVLDLFRELLNEGLEALKQKLK                                                                        67%                                              14  (SEQ ID NO: 14)                                                                        PVLDLFRELLNELLEALOQOLO                                                                        61%   70  96  80                                 15  (SEQ ID NO: 15)                                                                        PVLDLFRELWNELLEALKQKLK                                                                        60%   55  60  64  68                             16  (SEQ ID NO: 16)                                                                        PVLDLLRELLNELLEALKQKLK                                                                        59%                                              17  (SEQ ID NO: 17)                                                                        PVLELFKELLQELLEALKQKLK                                                                        59%                                              18  (SEQ ID NO: 18)                                                                        GVLDLFRELLNELLEALKQKLK                                                                        58%                                              19  (SEQ ID NO: 19)                                                                        pVLDLFRELLNEGLEALKQKLK                                                                        58%                                              20  (SEQ ID NO: 20)                                                                        PVLDLFREGLNELLEALKQKLK                                                                        57%                                              21  (SEQ ID NO: 21)                                                                        pVLDLFRELLNELLEALKQKLK                                                                        57%                                              22  (SEQ ID NO: 22)                                                                        PVLDLFRELLNELLEGLKQKLK                                                                        54%                                              23  (SEQ ID NO: 23)                                                                        PLLELFKELLQELLEALKQKLK                                                                        54%                                              24  (SEQ ID NO: 24)                                                                        PVLDLFRELLNELLEALQKKLK                                                                        53%                                              25  (SEQ ID NO: 25)                                                                        PVLDFFRELLNEXLEALKQKLK                                                                        51%   46  82      93                             26  (SEQ ID NO: 26)                                                                        PVLDLFRELLNELLELLKQKLK                                                                        47%                                              27  (SEQ ID NO: 27)                                                                        PVLDLFRELLNELZEALKQKLK                                                                        44%   72  92  82  81                             28  (SEQ ID NO: 28)                                                                        PVLDLFRELLNELWEALKQKLK                                                                        40%   82  98  90  81                             29  (SEQ ID NO: 29)                                                                        AVLDLFRELLNELLEALKQKLK                                                                        39%                                              30  (SEQ ID NO: 30)                                                                        PVLDLFRELLNELLEALKQKLK.sup.t                                                                  38%   85  90  98  90                             31  (SEQ ID NO: 31)                                                                        PVLDLFLELLNEXLEALKQKLK                                                                        34%   49  98      90                             32  (SEQ ID NO: 32)                                                                        XVLDLFRELLNELLEALKQKLK                                                                        33%                                              33  (SEQ ID NO: 33)                                                                        PVLDLFREKLNELLEALKQKLK                                                                        33%                                              34  (SEQ ID NO: 34)                                                                        PVLDZFRELLNELLEALKQKLK                                                                        32%   58  67  68  62                             35  (SEQ ID NO: 35)                                                                        PVLDWFRELLNELLEALKQKLK                                                                        31%   49  59  61                                                                    (sp)                                       36  (SEQ ID NO: 36)                                                                        PLLELLKELLQELLEALKQKLK                                                                        31%   95  100     95                             37  (SEQ ID NO: 37)                                                                        PVLDLFREWLNELLEALKQKLK                                                                        29%   65  75  76  73                             38  (SEQ ID NO: 38)                                                                        PVLDLFRELLNEXLEAWKQKLK                                                                        29%   25  49  21  49                             39  (SEQ ID NO: 39)                                                                        PVLDLFRELLEELLKALKKKLK                                                                        25%   66  69  68  72                             40  (SEQ ID NO: 40)                                                                        PVLDLFNELLRELLEALQKKLK                                                                        25%   66  84  79  77                             41  (SEQ ID NO: 41)                                                                        PVLDLWRELLNEXLEALKQKLK                                                                        25%   53  73  85  69                             42  (SEQ ID NO: 42)                                                                        PVLDEFREKLNEXWEALKQKLK                                                                        25%   15  74  27  76                             43  (SEQ ID NO: 43)                                                                        PVLDEFREKLWEXLEALKQKLK                                                                        25%                                              44  (SEQ ID NO: 44)                                                                        pvldefreklneXlealkqklk                                                                        25%   20  86                                     45  (SEQ ID NO: 45)                                                                        PVLDEFREKLNEXLEALKQKLK                                                                        24%   24  84  25  86                             46  (SEQ ID NO: 46)                                                                        PVLDLFREKLNEXLEALKQKLK                                                                        23%   30  86  58  85                             47  (SEQ ID NO: 47)                                                                        ˜VLDLFRELLNEGLEALKQKLK                                                                  23%                                              48  (SEQ ID NO: 48)                                                                        pvLDLFRELLNELLEALKQKLK                                                                        22%                                              49  (SEQ ID NO: 49)                                                                        PVLDLFRNLLEKLLEALEQKLK                                                                        22%   57  65  52  57                             50  (SEQ ID NO: 50)                                                                        PVLDLFRELLWEXLEALKQKLK                                                                        21%   68  84  89  76                             51  (SEQ ID NO: 51)                                                                        PVLDLFWELLNEXLEALKQKLK                                                                        20%   63  82  81  73                             52  (SEQ ID NO: 52)                                                                        PVWDEFREKLNEXLEALKQKLK                                                                        20%   sp  sp  sp                                 53  (SEQ ID NO: 53)                                                                        VVLDLFRELLNELLEALKQKLK                                                                        19%                                              54  (SEQ ID NO: 54)                                                                        PVLDLFRELLNEWLEALKQKLK                                                                        19%   76  71  84  78                             55  (SEQ ID NO: 55)                                                                        P˜˜˜LFRELLNELLEALKQKLK                                                      19%   38  72  78  75                             56  (SEQ ID NO: 56)                                                                        PVLDLFRELLNELLEALKQKKK                                                                        18%                                              57  (SEQ ID NO: 57)                                                                        PVLDLFRNLLEELLKALEQKLK                                                                        18%                                              58  (SEQ ID NO: 58)                                                                        PVLDEFREKLNEXLEALKQKL˜                                                                  18%                                              59  (SEQ ID NO: 59)                                                                        LVLDLFRELLNELLEALKQKLK                                                                        17%                                              60  (SEQ ID NO: 60)                                                                        PVLDLFRELLNELLEALKQ˜˜˜                                                      16%   39  83  66  84                             61  (SEQ ID NO: 61)                                                                        PVLDEFRWKLNEXLEALKQKLK                                                                        16%                                              62  (SEQ ID NO: 62)                                                                        PVLDEWREKLNEXLEALKQKLK                                                                        16%   15  85  43                                 63  (SEQ ID NO: 63)                                                                        PVLDFFREKLNEXLEALKQKLK                                                                        16%                                              64  (SEQ ID NO: 64)                                                                        PWLDEFREKLNEXLEALKQKLK                                                                        15%                                              65  (SEQ ID NO: 65)                                                                        ˜VLDEFREKLNEXLEALKQKLK                                                                  15%                                              66  (SEQ ID NO: 66)                                                                        PVLDLFRNLLEELLEALQKKLK                                                                        15%   64  82  66  70                             67  (SEQ ID NO: 67)                                                                        ˜VLDLFRELLNELLEALKQKLK                                                                  14%   81  90  84  94                             68  (SEQ ID NO: 68)                                                                        PVLDEFRELLKEXLEALKQKLK                                                                        14%                                              69  (SEQ ID NO: 69)                                                                        PVLDEFRKKLNEXLEALKQKLK                                                                        13%                                              70  (SEQ ID NO: 70)                                                                        PVLDEFRELLYEXLEALKQKLK                                                                        12%   27  78  33  66                             71  (SEQ ID NO: 71)                                                                        PVLDEFREKLNELXEALKQKLK                                                                        11%                                              72  (SEQ ID NO: 72)                                                                        PVLDLFRELLNEXLWALKQKLK                                                                        11%   sp  sp  sp                                 73  (SEQ ID NO: 73)                                                                        PVLDEFWEKLNEXLEALKQKLK                                                                        10%                                              74  (SEQ ID NO: 74)                                                                        PVLDKFREKLNEXLEALKQKLK                                                                        10%                                              75.sup.1/                                                                         (SEQ ID NO: 75)                                                                        PVLDEFREKLNEELEALKQKLK                                                                        10%   18  28  23  55                             76  (SEQ ID NO: 76)                                                                        PVLDEFRELLFEXLEALKQKLK                                                                         9%   41  88      66                             77  (SEQ ID NO: 77)                                                                        PVLDEFREKLNKXLEALKQKLK                                           78  (SEQ ID NO: 78)                                                                        PVLDEFRDKLNEXLEALKQKLK                                           79  (SEQ ID NO: 79)                                                                        PVLDEFRELLNELLEALKQKLK                                                                         9%                                              80  (SEQ ID NO: 80)                                                                        PVLDLFERLLNELLEALQKKLK                                                                         9%                                              81  (SEQ ID NO: 81)                                                                        PVLDEFREKLNWXLEALKQKLK                                           82  (SEQ ID NO: 82)                                                                        ˜˜LDEFREKLNEXLEALKQKLK                                                             8%                                              83  (SEQ ID NO: 83)                                                                        PVLDEFREKLNEXLEALWQKLK                                           84  (SEQ ID NO: 84)                                                                        PVLDEFREKLNELLEALKQKLK                                                                         7%                                              85  (SEQ ID NO: 85)                                                                        PVLDLFRELLNELLEALKQKLK                                                                         7%   58  61  64  69                             86  (SEQ ID NO: 86)                                                                        PVLELFERLLDELLNALQKKLK                                                                         7%                                              87  (SEQ ID NO: 87)                                                                        pllellkellqellealkqklk                                                                         7%   100 100     100                            88  (SEQ ID NO: 88)                                                                        PVLDKFRELLNEXLEALKQKLK                                                                         7%                                              89  (SEQ ID NO: 89)                                                                        PVLDEFREKLNEXLWALKQKLK                                                                         6%                                              90  (SEQ ID NO: 90)                                                                        ˜˜˜DEFREKLNEXLEALKQKLK                                                       6%                                              91  (SEQ ID NO: 91)                                                                        PVLDEFRELLNEXLEALKQKLK                                                                         6%   43  100     100                            92  (SEQ ID NO: 92)                                                                        PVLDEFRELYNEXLEALKQKLK                                                                         5%                                              93  (SEQ ID NO: 93)                                                                        PVLDEFREKLNEXLKALKQKLK                                                                         5%                                              94.sup.2/                                                                         (SEQ ID NO: 94)                                                                        PVLDEFREKLNEALEALKQKLK                                                                         5%   18  70  27  63                             95  (SEQ ID NO: 95)                                                                        PVLDLFRELLNLXLEALKQKLK                                                                         5%       sp  sp                                 96  (SEQ ID NO: 96)                                                                        pvldlfrellneXlealkqklk                                                                         5%   52  85  63  81                             97  (SEQ ID NO: 97)                                                                        PVLDLFRELLNELLE˜˜˜˜˜˜.ab                  out.             4%                                              98  (SEQ ID NO: 98)                                                                        PVLDLFRELLNEELEALKQKLK                                                                         2%                                              99  (SEQ ID NO: 99)                                                                        KLKQKLAELLENLLERFLDLVP                                                                         2%   72  88  80  80                            100  (SEQ ID NO: 100)                                                                       pvldlfrellnellealkqklk                                                                         2%   83  92      98                            101  (SEQ ID NO: 101)                                                                       PVLDLFRELLNWXLEALKQKLK                                                                         2%       sp  sp                                102  (SEQ ID NO: 102)                                                                       PVLDLFRELLNLXLEALKEKLK                                                                         2%   sp                                        103  (SEQ ID NO: 103)                                                                       PVLDEFRELLNEELEALKQKLK                                                                         1%                                             104  (SEQ ID NO: 104)                                                                       P˜˜˜˜˜˜˜LLNELLEALK                  QKLK            1%    21  49  29  55                            105  (SEQ ID NO: 105)                                                                       PAADAFREAANEAAEAAKQKAK                                                                         1%   29  28  32  65                            106  (SEQ ID NO: 106)                                                                       PVLDLFREKLNEELEALKQKLK                                                                         0%                                             107  (SEQ ID NO: 107)                                                                       klkqklaellenllerfldlvp                                                                         0%   sp  sp      77                            108  (SEQ ID NO: 108)                                                                       PVLDLFRWLLNEXLEALKQKLK                                                                         0%   28  55      54                             109.sup.3/                                                                        (SEQ ID NO: 109)                                                                       PVLDEFREKLNERLEALKQKLK                                                                         0%   19  45  23  58                            110  (SEQ ID NO: 110)                                                                       PVLDEFREKLNEXXEALKQKLK                                                                         0%                                             111  (SEQ ID NO: 111)                                                                       PVLDEFREKLWEXWEALKQKLK                                                                         0%                                             112  (SEQ ID NO: 112)                                                                       PVLDEFREKLNEXSEALKQKLK                                                                         0%                                             113  (SEQ ID NO: 113)                                                                       PVLDEFREKLNEPLEALKQKLK                                                                         0%    6  22                                    114  (SEQ ID NO: 114)                                                                       PVLDEFREKLNEXMEALKQKLK                                                                         0%                                             115  (SEQ ID NO: 115)                                                                       PKLDEFREKLNEXLEALKQKLK                                                                         0%                                             116  (SEQ ID NO: 116)                                                                       PHLDEFREKLNEXLEALKQKLK                                                                         0%                                             117  (SEQ ID NO: 117)                                                                       PELDEFREKLNEXLEALKQKLK                                                                         0%                                             118  (SEQ ID NO: 118)                                                                       PVLDEFREKLNEXLEALEQKLK                                                                         0%                                             119  (SEQ ID NO: 119)                                                                       PVLDEFREKLNEELEAXKQKLK                                                                         0%                                             120  (SEQ ID NO: 120)                                                                       PVLDEFREKLNEELEXLKQKLK                                                                         0%                                             121  (SEQ ID NO: 121)                                                                       PVLDEFREKLNEELEALWQKLK                                                                         0%                                             122  (SEQ ID NO: 122)                                                                       PVLDEFREKLNEELEWLKQKLK                                                                         0%                                             123  (SEQ ID NO: 123)                                                                       QVLDLFRELLNELLEALKQKLK                                          124  (SEQ ID NO: 124)                                                                       PVLDLFOELLNELLEALOQOLO                                          125  (SEQ ID NO: 125)                                                                       NVLDLFRELLNELLEALKQKLK                                          126  (SEQ ID NO: 126)                                                                       PVLDLFRELLNELGEALKQKLK                                          127  (SEQ ID NO: 127)                                                                       PVLDLFRELLNELLELLKQKLK                                                                        47%                                             128  (SEQ ID NO: 128)                                                                       PVLDLFRELLNELLEFLKQKLK                                          129  (SEQ ID NO: 129)                                                                       PVLELFNDLLRELLEALQKKLK                                          130  (SEQ ID NO: 130)                                                                       PVLELFNDLLRELLEALKQKLK                                          131  (SEQ ID NO: 131)                                                                       PVLELFKELLNELLDALRQKLK                                          132  (SEQ ID NO: 132)                                                                       PVLDLFRELLENLLEALQKKLK                                          133  (SEQ ID NO: 133)                                                                       PVLELFERLLEDLLQALNKKLK                                          134  (SEQ ID NO: 134)                                                                       PVLELFERLLEDLLKALNQKLK                                          135  (SEQ ID NO: 135)                                                                       DVLDLFRELLNELLEALKQKLK                                          136  (SEQ ID NO: 136)                                                                       PALELFKDLLQELLEALKQKLK                                          137  (SEQ ID NO: 137)                                                                       PVLDLFRELLNEGLEAZKQKLK                                          138  (SEQ ID NO: 138)                                                                       PVLDLFRELLNEGLEWLKQKLK                                          139  (SEQ ID NO: 139)                                                                       PVLDLFRELWNEGLEALKQKLK                                          140  (SEQ ID NO: 140)                                                                       PVLDLFRELLNEGLEALOQOLO                                          141  (SEQ ID NO: 141)                                                                       PVLDFFRELLNEGLEALKQKLK                                          142  (SEQ ID NO: 142)                                                                       PVLELFRELLNEGLEALKQKLK                                          143  (SEQ ID NO: 143)                                                                       PVLDLFRELLNEGLEALKQKLK*                                         144  (SEQ ID NO: 144)                                                                       PVLELFENLLERLLDALQKKLK                                                                        111%  89  88      95                            145  (SEQ ID NO: 145)                                                                       GVLELFENLLERLLDALQKKLK                                                                        100%  55  51      58                            146  (SEQ ID NO: 146)                                                                       PVLELFENLLERLLDALQKKLK                                                                        86%   97  100 100 95                            147  (SEQ ID NO: 147)                                                                       PVLELFENLLERLFDALQKKLK                                                                        76%                                             148  (SEQ ID NO: 148)                                                                       PVLELFENLLERLGDALQKKLK                                                                        75%   10  76  23  80                            149  (SEQ ID NO: 149)                                                                       PVLELFENLWERLLDALQKKLK                                                                        63%   28  54      47                            150  (SEQ ID NO: 150)                                                                       PLLELFENLLERLLDALQKKLK                                                                        57%                                             151  (SEQ ID NO: 151)                                                                       PVLELFENLGERLLDALQKKLK                                                                        55%                                             152  (SEQ ID NO: 152)                                                                       PVFELFENLLERLLDALQKKLK                                                                        50%                                             153  (SEQ ID NO: 153)                                                                       AVLELFENLLERLLDALQKKLK                                                                        49%                                             154  (SEQ ID NO: 154)                                                                       PVLELFENLLERGLDALQKKLK                                                                        39%   13  76  25  80                            155  (SEQ ID NO: 155)                                                                       PVLELFLNLWERLLDALQKKLK                                                                        38%                                             156  (SEQ ID NO: 156)                                                                       PVLELFLNLLERLLDALQKKLK                                                                        35%                                             157  (SEQ ID NO: 157)                                                                       PVLEFFENLLERLLDALQKKLK                                                                        30%                                             158  (SEQ ID NO: 158)                                                                       PVLELFLNLLERLLDWLQKKLK                                                                        30%                                             159  (SEQ ID NO: 159)                                                                       PVLDLFENLLERLLDALQKKLK                                                                        28%                                             160  (SEQ ID NO: 160)                                                                       PVLELFENLLERLLDWLQKKLK                                                                        28%   65  73  75  61                            161  (SEQ ID NO: 161)                                                                       PVLELFENLLERLLEALQKKLK                                                                        27%                                             162  (SEQ ID NO: 162)                                                                       PVLELFENWLERLLDALQKKLK                                                                        27%   68  83  81                                163  (SEQ ID NO: 163)                                                                       PVLELFENLLERLWDALQKKLK                                                                        26%   27  53      55                            164  (SEQ ID NO: 164)                                                                       PVLELFENLLERLLDAWQKKLK                                                                        24%   37  66  51  61                            165  (SEQ ID NO: 165)                                                                       PVLELFENLLERLLDLLQKKLK                                                                        23%                                             166  (SEQ ID NO: 166)                                                                       PVLELFLNLLEKLLDALQKKLK                                                                        22%                                             167  (SEQ ID NO: 167)                                                                       PVLELFENGLERLLDALQKKLK                                                                        18%                                             168  (SEQ ID NO: 168)                                                                       PVLELFEQLLEKLLDALQKKLK                                                                        17%                                             169  (SEQ ID NO: 169)                                                                       PVLELFENLLEKLLDALQKKLK                                                                        17%                                             170  (SEQ ID NO: 170)                                                                       PVLELFENLLEOLLDALQOOLO                                                                        17%                                             171  (SEQ ID NO: 171)                                                                       PVLELFENLLEKLLDLLQKKLK                                                                        16%                                             172  (SEQ ID NO: 172)                                                                       PVLELFLNLLERLGDALQKKLK                                                                        16%                                             173  (SEQ ID NO: 173)                                                                       PVLDLFDNLLDRLLDLLNKKLK                                                                        15%                                             174  (SEQ ID NO: 174)                                                                       pvlelfenllerlldalqkklk                                                                        13%                                             175  (SEQ ID NO: 175)                                                                       PVLELFENLLERLLELLNKKLK                                                                        13%                                             176  (SEQ ID NO: 176)                                                                       PVLELWENLLERLLDALQKKLK                                                                        11%                                             177  (SEQ ID NO: 177)                                                                       GVLELFLNLLERLLDALQKKLK                                                                        10%                                             178  (SEQ ID NO: 178)                                                                       PVLELFDNLLEKLLEALQKKLR                                                                         9%                                             179  (SEQ ID NO: 179)                                                                       PVLELFDNLLERLLDALQKKLK                                                                         8%                                             180  (SEQ ID NO: 180)                                                                       PVLELFDNLLDKLLDALQKKLR                                                                         8%                                             181  (SEQ ID NO: 181)                                                                       PVLELFENLLERWLDALQKKLK                                                                         8%                                             182  (SEQ ID NO: 182)                                                                       PVLELFENLLEKLLEALQKKLK                                                                         7%                                             183  (SEQ ID NO: 183)                                                                       PLLELFENLLEKLLDALQKKLK                                                                         6%                                             184  (SEQ ID NO: 184)                                                                       PVLELFLNLLERLLDAWQKKLK                                                                         4%                                             185  (SEQ ID NO: 185)                                                                       PVLELFENLLERLLDALQOOLO                                                                         3%                                             186  (SEQ ID NO: 186)                                                                       PVLELFEQLLERLLDALQKKLK                                          187  (SEQ ID NO: 187)                                                                       PVLELFENLLERLLDALNKKLK                                          188  (SEQ ID NO: 188)                                                                       PVLELFENLLDRLLDALQKKLK                                          189  (SEQ ID NO: 189)                                                                       DVLELFENLLERLLDALQKKLK                                          190  (SEQ ID NO: 190)                                                                       PVLEFWDNLLDKLLDALQKKLR                                          191  (SEQ ID NO: 191)                                                                       PVLDLLRELLEELKQKLK*                                                                           100%                                            192  (SEQ ID NO: 192)                                                                       PVLDLFKELLEELKQKLK*                                                                           100%  36          56                            193  (SEQ ID NO: 193)                                                                       PVLDLFRELLEELKQKLK*                                                                           96%   34  88  87  87                            194  (SEQ ID NO: 194)                                                                       PVLELFRELLEELKQKLK*                                                                           88%   38  93      93                            195  (SEQ ID NO: 195)                                                                       PVLELFKELLEELKQKLK*                                                                           87%                                             196  (SEQ ID NO: 196)                                                                       PVLDLFRELLEELKNKLK*                                                                           81%                                             197  (SEQ ID NO: 197)                                                                       PLLDLFRELLEELKQKLK*                                                                           81%   43  70      69                            198  (SEQ ID NO: 198)                                                                       GVLDLFRELLEELKQKLK*                                                                           80%                                             199  (SEQ ID NO: 199)                                                                       PVLDLFRELWEELKQKLK*                                                                           76%   35  77  80  79                            200  (SEQ ID NO: 200)                                                                       NVLDLFRELLEELKQKLK*                                                                           75%                                             201  (SEQ ID NO: 201)                                                                       PLLDLFKELLEELKQKLK*                                                                           74%                                             202  (SEQ ID NO: 202)                                                                       PALELFKDLLEELRQKLR*                                                                           70%                                             203  (SEQ ID NO: 203)                                                                       AVLDLFRELLEELKQKLK*                                                                           66%                                             204  (SEQ ID NO: 204)                                                                       PVLDFFRELLEELKQKLK*                                                                           63%                                             205  (SEQ ID NO: 205)                                                                       PVLDLFREWLEELKQKLK*                                                                           60%                                             206  (SEQ ID NO: 206)                                                                       PLLELLKELLEELKQKLK*                                                                           57%                                             207  (SEQ ID NO: 207)                                                                       PVLELLKELLEELKQKLK*                                                                           50%                                             208  (SEQ ID NO: 208)                                                                       PALELFKDLLEELRQRLK*                                                                           48%                                             209  (SEQ ID NO: 209)                                                                       PVLDLFRELLNELLQKLK                                                                            47%   54  71  67  62                            210  (SEQ ID NO: 210)                                                                       PVLDLFRELLEELKQKLK                                                                            46%   20  63  37  53                            211  (SEQ ID NO: 211)                                                                       PVLDLFRELLEELOQOLO*                                                                           45%                                             212  (SEQ ID NO: 212)                                                                       PVLDLFOELLEELOQOLK*                                                                           43%                                             213  (SEQ ID NO: 213)                                                                       PALELFKDLLEEFRQRLK*                                                                           42%                                             214  (SEQ ID NO: 214)                                                                       pVLDLFRELLEELKQKLK*                                                                           39%                                             215  (SEQ ID NO: 215)                                                                       PVLDLFRELLEEWKQKLK*                                                                           38%   28  63  53  68                            216  (SEQ ID NO: 216)                                                                       PVLELFKELLEELKQKLK                                                                            35%                                             217  (SEQ ID NO: 217)                                                                       PVLDLFRELLELLKQKLK                                                                            30%   52  78  76  70                            218  (SEQ ID NO: 218)                                                                       PVLDLFRELLNELLQKLK*                                                                           29%                                             219  (SEQ ID NO: 219)                                                                       PVLDLFRELLNELWQKLK                                                                            24%                                             220  (SEQ ID NO: 220)                                                                       PVLDLFRELLEELQKKLK                                                                            22%   27  64  54  64                            221  (SEQ ID NO: 221)                                                                       DVLDLFRELLEELKQKLK*                                                                           12%                                             222  (SEQ ID NO: 222)                                                                       PVLDAFRELLEALLQLKK                                                                             8%                                             223  (SEQ ID NO: 223)                                                                       PVLDAFRELLEALAQLKK                                                                             8%   21  56  23  51                            224  (SEQ ID NO: 224)                                                                       PVLDLFREGWEELKQKLK                                                                             8%                                             225  (SEQ ID NO: 225)                                                                       PVLDAFRELAEALAQLKK                                                                             1%                                             226  (SEQ ID NO: 226)                                                                       PVLDAFRELGEALLQLKK                                                                             1%                                             227  (SEQ ID NO: 227)                                                                       PVLDLFRELGEELKQKLK*                                                                            0%                                             228  (SEQ ID NO: 228)                                                                       PVLDLFREGLEELKQKLK*                                                                            0%                                             229  (SEQ ID NO: 229)                                                                       PVLDLFRELLEEGKQKLK*                                                                            0%                                             230  (SEQ ID NO: 230)                                                                       PVLELFERLLEDLQKKLK                                              231  (SEQ ID NO: 231)                                                                       PVLDLFRELLEKLEQKLK                                              232  (SEQ ID NO: 232)                                                                       PLLELFKELLEELKQKLK*                                              237.sup.4/                                                                        (SEQ ID NO: 237)                                                                       LDDLLQKWAEAFNQLLKK                                                                            11%   30  66  45  --                             238.sup.5/                                                                        (SEQ ID NO: 238)                                                                       EWLKAFYEKVLEKLKELF*                                                                           19%   49  72  60  58                             239.sup.6/                                                                        (SEQ ID NO: 239)                                                                       EWLEAFYKKVLEKLKELF*                                                                           11%   44  49      sp                            240  (SEQ ID NO: 240)                                                                       DWLKAFYDKVAEKLKEAF*                                                                           10%   16  68  59  57                            241  (SEQ ID NO: 241)                                                                       DWFKAFYDKVFEKFKEFF                                                                             8%                                              242.sup.7/                                                                        (SEQ ID NO: 242)                                                                       GIKKFLGSIWKFIKAFVG                                                                             7%                                             243  (SEQ ID NO: 243)                                                                       DWFKAFYDKVAEKFKEAF                                                                             5%   10  64  50                                 244.sup.8/                                                                        (SEQ ID NO: 244)                                                                       DWLKAFYDKVAEKLKEAF                                                                             5%    9  40  13  48                            245  (SEQ ID NO: 245)                                                                       DWLKAFYDKVFEKFKEFF                                                                             4%   38  77  70  sp                             246.sup.9/                                                                        (SEQ ID NO: 246)                                                                       EWLEAFYKKVLEKLKELF                                                                             4%   18  44  47                                247  (SEQ ID NO: 247)                                                                       DWFKAFYDKFFEKFKEFF                                                                             3%                                              248.sup.10/                                                                       (SEQ ID NO: 248)                                                                       EWLKAFYEKVLEKLKELF                                                                             3%   18  45  13                                 249.sup.11/                                                                       (SEQ ID NO: 249)                                                                       EWLKAEYEKVEEKLKELF*                                              250.sup.12/                                                                       (SEQ ID NO: 250)                                                                       EWLKAEYEKVLEKLKELF*                                              251.sup.13/                                                                       (SEQ ID NO: 251)                                                                       EWLKAFYKKVLEKLKELF*                                             252  (SEQ ID NO: 252)                                                                       PVLDLFRELLEQKLK*                                                253  (SEQ ID NO: 2S3)                                                                       PVLDLFRELLEELKQK*                                               254  (SEQ ID NO: 254)                                                                       PVLDLFRELLEKLKQK*                                               255  (SEQ ID NO: 255)                                                                       PVLDLFRELLEKLQK*                                                256  (SEQ ID NO: 256)                                                                       PVLDLFRELLEALKQK*                                               257  (SEQ ID NO: 257)                                                                       PVLDLFENLLERLKQK*                                               258  (SEQ ID NO: 258)                                                                       PVLDLFRELLNELKQK*                                               __________________________________________________________________________     .sup.1/ Segrest's Consensus 22mer peptide (Anantharamaiah et al., 1990,       Arteriosclerosis 10(1):95-105).                                               .sup.2/ [A.sup.13 ] Consensus 22mer peptide (Anantharamaiah et al., 1990,     Arteriosclerosis 10(1):95-105).                                               .sup.3/ [R.sup.13 ] Consensus 22mer peptide (Anantharamaiah et al., 1990,     Arteriosclerosis 10(1):95-105).                                               .sup.4/ ID3 peptide (Labeur et al., 1997, Arteriosclerosis, Thrombosis an     Vascular Biology 17(3):580-588).                                              .sup.5/ Ac18AMOD  C(O)NH.sub.2 peptide (Epand et al., 1987, J. Biol. Chem     262(19):9389-9396).                                                           .sup.6/ Ac18AM4  C(O)NH.sub.2 peptide (Brasseur, 1993, Biochim. Biophys.      Acta 1170:1-7).                                                               .sup.7/ 18L peptide (Segrest et al., 1990, Proteins: Structure,Function       and Genetics 8:103-117).                                                      .sup.8/ 18A peptide (Anantharamaiah et al., 1985, J. Biol. Chem.              260(18):10248-10255).                                                         .sup.9/ 18AM4 peptide (Rosseneu et al., WO93/25581; Corijn et al. 1993,       Biochim. Biophys. Acta 1170:8-16).                                            .sup.10/ [Glu.sup.1,8 ; Leu.sup.5,11,17] 18A peptide (Epand et al., 1987,     J. Biol. Chem. 262(19):9389-9396).                                            .sup.11/ Ac18AM3  C(O)NH.sub.2 (Rosseneu et al., WO93/25581).                 .sup.12/ Ac18AM2  C(O)NH.sub.2 (Rosseneu et al., WO93/25581).                 .sup.13/ Ac18AM1  C(O)NH.sub.2 (Rosseneu et al., WO93/25581).            

In TABLE IX, * indicates peptides that are N-terminal acetylated andC-terminal amidated; ↑ indicates peptides that are N-terminaldansylated; sp indicates peptides that exhibited solubility problemsunder the experimental conditions; X is Aib; Z is Nal; O is Orn; He (%)designates percent helicity; mics designates micelles; and ˜ indicatesdeleted amino acids.

9. EXAMPLE Pharmacokinetics of the ApoA-I Agonists

The following experiments can be used to demonstrate that the ApoA-Iagonists are stable in the circulation and associate with the HDLcomponent of plasma.

9.1. Synthesis of Radiolabelled Peptides

Radiolabelled peptides are synthesized by coupling a ¹⁴ C-labeled aminoacid as the N-terminal amino acid. The synthesis is carried outaccording to L. Lapatsanis, Synthesis, 1983, 671-173. Briefly, 250 μM ofunlabeled N-terminal amino acid is dissolved in 225 μl of a 9% Na₂ CO₃solution and added to a solution (9% Na₂ CO₃) of 9.25 MBq (250 μM) ¹⁴C-labeled N-terminal amino acid. The liquid is cooled down to 0° C.,mixed with 600 μM (202 mg) 9-fluorenylmethyl-N-succinimidylcarbonate(Fmoc-OSu) in 0.75 ml DMF and shaken at room temperature for 4 hr.Thereafter the mixture is extracted with Diethylether (2×5 ml) andchloroform (1×5 ml), the remaining aqueous phase is acidified with 30%HCl and extracted with chloroform (5×8 ml). The organic phase is driedover Na₂ SO₄, filtered off and the volume was reduced under nitrogenflow to 5 ml. The purity is estimated by TLC (CHCl₃ :MeOH:Hac, 9:1:0.1v/v/v, stationary phase RPTLC silicagel 60, Merck, Germany).

The chloroform solution containing ¹⁴ C-labeled Fmoc amino acid is useddirectly for peptide synthesis. A peptide resin containing amino acids2-22 is synthesized automatically as described in Section 6. Thesequence of the peptide is determined by Edman degradation. The couplingis performed as described in Section 6.1.

9.2. Pharmacokinetics in Mice

In each experiment, 2.5 mg/kg radiolabelled peptide is injectedintraperitoneally into mice which are fed normal mouse chow or theatherogenic Thomas-Harcroft modified diet (resulting in severelyelevated VLDL and IDL cholesterol). Blood samples are taken at multipletime intervals for assessment of radioactivity in plasma.

9.3. Stability in Human Serum

The stability of the ApoA-I agonists of the invention in human serum isdemonstrated as described below.

9.3.1. Experimental Methods

100 μg of ¹⁴ C-labeled peptide (prepared as described in Section 9.1,supra), is mixed with 2 ml of fresh human plasma (at 37° C.) anddelipidated either immediately (control sample) or after 8 days ofincubation at 37° C. (test sample). Delipidation is carried out byextracting the lipids with an equal volume of 2:1 (v/v)chloroform:methanol.

The samples are loaded onto a reverse-phase C₁₈ HPLC column and elutedwith a linear gradient (25-58% over 33 min.) of acetonitrile (containing0.1% TFA). Elution profiles are followed by absorbance (220 nm) andradioactivity.

9.4. Formation of Pre-β Like Particles

The ability of the ApoA-I agonists of the invention to form pre-β-likeparticles is demonstrated as described below.

9.4.1. Experimental Method

Human HDL is isolated by KBr density ultra centrifugation at densityd=1.21 g/ml to obtain top fraction followed by Superose 6 gel filtrationchromatography to separate HDL from other lipoproteins. Isolated HDL isadjusted to a final concentration of 1.0 mg/ml with physiological salinebased on protein content determined by Bradford protein assay. Analiquot of 300 μl is removed from the isolated HDL preparation andincubated with 100 μl ¹⁴ C-labeled peptide for two hours at 37° C. Fiveseparate incubations are analyzed including a blank containing 100 μlphysiological saline and four dilutions of ¹⁴ C-labeled peptide: (i)0.20 μg/μl peptide:HDL, ratio=1:15; (ii) 0.30 μg/μl peptide:HDL,ratio=1:10; (iii) 0.60 μg/μl peptide:HDL, ratio=1:5; and (iv) 1.00 μg/μlpeptide:HDL, ratio=1:3. Following the two hour incubation, a 200 μlaliquot of the sample (total volume=400 μl) is loaded onto a Superose 6gel filtration column for lipoprotein separation and analysis, and 100μl is used to determine total radioactivity loaded onto the column.

9.5. Association of Apo-A-I Agonists with Human Lipoproteins 9.5.1.Experimental Methods

The ability of the ApoA-I agonists of the invention to associate withhuman lipoprotein fractions is determined by incubating ¹⁴ C-labeledpeptide with each lipoprotein class (HDL, LDL and VLDL) and a mixture ofthe different lipoprotein classes.

HDL, LDL and VLDL are isolated by KBr density gradientultracentrifugation at d=1.21 g/ml and purified by FPLC on a Superose 6Bcolumn size exclusion column (chromatography is carried out at a flowrate of 0.7 ml/min. and a running buffer of 10 mM Tris (pH 8), 115 mMNaCl, 2 mM EDTA and 0.01% NaN₃). ¹⁴ C-labeled peptide is incubated withHDL, LDL and VLDL at a peptide:phospholipid ratio of 1:5 (mass ratio)for 2 h at 37° C. The required amount of lipoprotein (volumes based onamount needed to yield 1000 μg) is mixed with 0.2ml of peptide stocksolution (1 mg/ml) and the solution is brought up to 2.2 ml using 0.9%of NaCl.

After incubating for 2 hr. at 37° C., an aliquot (0.1 ml) is removed forliquid scintillation counting to determine the total radioactivity, thedensity of the remaining incubation mixture is adjusted to 1.21 g/mlwith KBr, and the samples are centrifuged at 100,000 rpm (300,000 g) for24 hours at 4° C. in a TLA 100.3 rotor using a Beckman tabletopultracentrifuge. The resulting supernatant is fractionated by removing0.3 ml aliquots from the top of each sample for a total of 5 fractions,and 0.05 ml of each fraction is used for liquid scintillation counting.The top two fractions contain the floating lipoproteins, the otherfractions (3-5) correspond to proteins/peptides in solution.

9.6. The ApoA-I Agonists of the Invention Selectively Bind HDL Lipids inHuman Plasma 9.6.1. Experimental Method

To demonstrate that the ApoA-I agonists of the invention selectivelybind HDL proteins in human plasma, 2 ml of human plasma is incubatedwith 20, 40, 60, 80, and 100 μg of ¹⁴ C-labeled peptide for 2 hr. at 37°C. The lipoproteins are separated by adjusting the density to 1.21 g/mland centrifugation in a TLA 100.3 rotor at 100,000 rpm (300,000 g) for36 hr. at 4° C. The top 900 μl (in 300 μl fractions) is taken foranalysis. 50 μl from each 300 μl fraction is counted for radioactivityand 200 μl from each fraction is analyzed by FPLC (Superose 6/Superose12 combination column).

10. EXAMPLE The ApoA-I Agonists Promote Cholesterol Efflux

To demonstrate that the ApoA-I agonists of the invention promotecholesterol efflux, HepG2 hepatoma cells are plated into 6-well culturedishes and grown to confluence. Cells are labeled with ³ H-cholesterolby drying the cholesterol, then adding 1% bovine serum albumin (BSA) inphosphate buffered saline (PBS), sonicating the solution, and adding 0.2ml of this labeling solution and 1.8 ml growth medium to the cells, sothat each well contains 2 μCi of radioactivity. Cells are incubated for24 hr. with the labeling medium.

Peptide (or protein):DMPC complexes are prepared at a 1:2 peptide (orprotein):DMPC ratio (w:w). To prepare the complexes, peptide or nativehuman ApoA-I protein is added to a DMPC solution in PBS and incubated atroom temperature overnight, by which time the solution will clarify.Peptide or protein concentration in the final solution is about 1 mg/ml.

Labeling media is removed from the cells and the cells are washed withPBS prior to addition of complexes. 1.6 ml of growth medium is added toeach well, followed by peptide (or protein):DMPC complex and sufficientPBS to bring the final volume to 2 ml per well. The final peptide orApoA-I concentrations are about 1, 2.5, 5, 7.5 and 25 μg/ml medium.After 24 hours of incubation at 37° C., the medium is removed, and thecells are washed with 2 ml of 1% BSA/PBS, followed by 2 washes with 2 mleach of PBS. The amount of ³ H-cholesterol effluxed into the medium isdetermined by liquid scintillation counting.

11. EXAMPLE Use of the ApoA-I Agonists in Animal Model Systems

The efficacy of the ApoA-I agonists of the invention can be demonstratedin rabbits using the protocols below.

11.1. Preparation of the Phospholipid/Peptide Complexes

Small discoidal particles consisting of phospholipid (DPPC) and peptideare prepared following the cholate dialysis method. The phospholipid isdissolved in chloroform and dried under a stream of nitrogen. Thepeptide is dissolved in buffer (saline) at a concentration of 1-2 mg/ml.The lipid film is redissolved in buffer containing cholate (43° C.) andthe peptide solution is added at a 3:1 phospholipid/peptide ratio. Themixture is incubated overnight at 43° C. and then dialyzed at 43° C. (24hr.), room temperature (24 hr.) and 4° C. (24 hr.), with three changesof buffer (large volumes) at temperature point. The complexes are filtersterilized (0.22μ) for injection and storage at 4° C.

11.2. Isolation and Characterization of the Peptide/PhospholipidParticles

The particles are separated on a gel filtration column (Superose 6 HR).The position of the peak containing the particles is identified bymeasuring the phospholipid concentration in each fraction. From theelution volume, the Stokes radius can be determined. The concentrationof peptide in the complex is determined by determining the phenylalaninecontent (by HPLC) following a 16 hr. acid hydrolysis.

11.3. Injection in the Rabbit

Male New Zealand White rabbits (2.5-3 kg) are injected intravenouslywith a dose of phospholipid/peptide complex (5-10 mg/kg bodyweightpeptide or 10 mg/kg bodyweight ApoA-I (control)), expressed as peptideor protein content) in a single bolus injection not exceeding 10-15 ml.The animals are slightly sedated before the manipulations. Blood samples(collected on EDTA) are taken before and 5, 15, 30, 60, 240 and 1440minutes after injection. The hematocrit (Hct) is determined for eachsample. Samples are aliquoted and stored at -20° C. before analysis.

11.4. Analysis of the Rabbit Sera

Plasma Lipids

The total plasma cholesterol, plasma triglycerides and plasmaphospholipids is determined enzymatically using commercially availableassays according to the manufacturer's protocols (Boehringer Mannheim,Mannheim, Germany and Biomerieux, 69280, Marcy-l'etoile, France).

Lipoprotein Profiles

The plasma lipoprotein profiles of the fractions obtained after theseparation of the plasma into its lipoprotein fractions is determined byspinning in a sucrose density gradient. The fractions are collected andin each individual fraction the phospholipid and cholesterol content ismeasured enzymatically.

12. EXAMPLE Preparation of Peptide-Lipid Complex by Co-LyophilizationApproach

The following protocol was utilized to prepare peptide-lipid complexes.

One mg of peptide 210 (SEQ ID NO:210) was dissolved in 250 μl HPLC grademethanol (Perkin Elmer) in a one ml clear glass vial with cap (Waters#WAT025054). Dissolving of the peptide was aided by occasional vortexingover a period of 10 minutes at room temperature. To this mixture analiquot containing 3 mg dipalmitoyl-phosphatidylcholine (DPPC; AvantiPolar Lipids, 99% Purity, product #850355) from a 100 mg/ml stocksolution in methanol was added. The volume of the mixture was brought to400 μl by addition of methanol, and the mixture was further vortexedintermittently for a period of 10 minutes at room temperature. To thetube 200 μl of xylene (Sigma-Aldrich 99% pure, HPLC-grade) was added andthe tubes were vortexed for 10 seconds. Two small holes were punchedinto the top of the tube with a 20 gauge syringe needle, the tube wasfrozen for 15 seconds in liquid nitrogen, and the tube was lyophilizedovernight under vacuum. To the tube 200 mls of 0.9% NaCl solution wasadded. The tube was vortexed for 20 seconds. At this time the solutionin the tube was milky in appearance. The tube was then incubated in awater bath for 30 minutes at 41° C. The solution became clear (i.e.,similar to water in appearance) after a few minutes of incubation at 41°C.

12.1. Characterization of Complexes by Superose 6 Gel FiltrationChromatography

Peptide-phospholipid complexes containing peptide 210 (SEQ ID NO:210)were prepared by colyophilization as described above. The preparationcontained 1 mg peptide and 3 mg DPPC by weight. After reconstituting thecomplexes in 200 μl of 0.9% NaCl, 20 μl (containing 100 μg peptide 210)of the complexes were applied to a Pharmacia Superose 6 column using0.9% NaCl as the liquid phase at a flow rate of 0.5 ml/minute. Thechromatography was monitored by absorbance or scattering of light ofwavelength 280 nm. One ml fractions were collected. Aliquots containing20 μl of the fractions were assayed for phospholipid content using thebioMerieux Phospholipides Enzymatique PAP 150 kit (#61491) according tothe instructions supplied by the manufacturer. The vast majority of bothphospholipid and UV absorbance were recovered together in a fewfractions with peaks at approximately 15.1 mls. This elution volumecorresponds to a Stokes' diameter of 106 Angstroms.

For comparison, a separate chromatogram of 20 μl of human HDL₂ was rununder the same conditions and using the same column as the peptide 210(SEQ ID NO:210) complexes. The HDL₂ was prepared as follows: 300 mlsfrozen human plasma (Mannheim Blutspendzentrale #1185190) was thawed,adjusted to density 1.25 with solid potassium bromide, and centrifuged45 hours at 40,000 RPM using a Ti45 rotor (Beckman) at 20° C. Thefloating layer was collected, dialyzed against distilled water, adjustedto density 1.07 with solid potassium bromide, and centrifuged asdescribed above for 70 hours. The bottom layer (at a level of one cmabove the tube bottom) was collected, brought to 0.01% sodium azide, andstored at 4° C. for 4 days until chromatography. The column eluate wasmonitored by absorbance or scattering of light of wavelength 254 nm. Aseries of proteins of known molecular weight and Stokes' diameter wereused as standards to calibrate the column for the calculation of Stokes'diameters of the particles (Pharmacia Gel Filtration Calibration KitInstruction Manual, Pharmacia Laboratory Separation, Piscataway, N.J.,revised April, 1985). The HDL₂ eluted with a retention volume of 14.8mls, corresponding to a Stokes' diameter of 108 nm.

13. EXAMPLE Preparation of Antibodies

To prepare antibodies to the ApoA-I agonists of the invention, peptideis conjugated to keyhole limpet hemocyanine (KLH; 1 mg peptide to 10 mgKLH). The KLH conjugate (LMG) is suspended in complete Freund's adjuvantand injected into rabbits at time 0, and boosted with 0.25 mg KLHconjugate at 4 weeks and again at 5 weeks. Pre-bleeds and six weekpost-bleeds are tested for antibody titer against authentic antigen byELISA.

The production bleeds are pooled from 2 rabbits each. Antibodiesdirected exclusively against the peptide antigens are isolated asfollows:

1. Free peptide is attached to cyanogen bromide activated Sepharose 4B(Pharmacia) according to the manufacturer's protocol.

2. The antisera is preabsorbed on a column of irrelevant peptides and oncolumns of irrelevant human and mouse serum proteins.

3. The pre-absorbed antisera is passed through the corresponding peptidecolumn (see point 1).

4. The columns are washed with 0.1 M borate buffered saline (pH 8.2) andthe bound antibodies are eluted using a low pH gradient step from pH 4.0to pH 3.0 to pH 2.0 (0.1 M glycine buffer) and finally with 0.1 M HCl.

5. The eluted material is neutralized with excess borate saline,concentrated by ultrafiltration (Amicon, YM30) and dialyzed againstborate saline.

6. The protein concentration is determined by absorbance at 280 nm.

The resulting antibodies are tested for species specificity usingpurified human ApoA-I or purified mouse ApoA-I in a direct ELISA bindingassay.

The invention is not to be limited in scope by the specific embodimentsdescribed which are intended as single illustrations of individualaspects of the invention, and functionally equivalent methods andcomponents are within the scope of the invention. Indeed variousmodifications of the invention, in addition to those shown and describedherein will become apparent to those skilled in the art from theforegoing description and accompanying drawings. Such modifications areintended to fall within the scope of the appended claims.

All references cited herein are incorporated herein by reference for allpurposes.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 258                                           - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 16                                                    #Xaa = NaphthylalanineFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Xaa         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 23 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 - Gly Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys Lys                                                             20                                                                - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Trp         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 23 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = D-Pro) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 - Xaa Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys Lys                                                             20                                                                - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 - Pro Val Leu Asp Leu Phe Lys Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 - Pro Val Leu Asp Leu Phe Arg Glu Leu Gly As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 17                                                    #Xaa = NaphthylalanineFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Xaa Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:11:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                - Pro Val Leu Asp Leu Phe Lys Glu Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:12:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Gly Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:13:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                - Gly Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:14:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 18                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 20                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 22                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Xaa Gln Xaa Leu Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:15:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Trp As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:16:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                - Pro Val Leu Asp Leu Leu Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:17:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                - Pro Val Leu Glu Leu Phe Lys Glu Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:18:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                - Gly Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:19:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = D-Pro) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                - Xaa Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:20:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                - Pro Val Leu Asp Leu Phe Arg Glu Gly Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:21:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = D-Pro) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                - Xaa Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:22:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Gly         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:23:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                - Pro Leu Leu Glu Leu Phe Lys Glu Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:24:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:25:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                - Pro Val Leu Asp Phe Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:26:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Leu         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:27:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 14                                                    #Xaa = NaphthylalanineFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Xaa Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:28:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Trp Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:29:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                - Ala Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:30:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #N-terminal dansylated peptideN:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:31:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                - Pro Val Leu Asp Leu Phe Leu Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:32:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                - Xaa Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:33:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                - Pro Val Leu Asp Leu Phe Arg Glu Lys Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:34:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 5                                                     #Xaa = NaphthylalanineFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                - Pro Val Leu Asp Xaa Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:35:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                - Pro Val Leu Asp Trp Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:36:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                - Pro Leu Leu Glu Leu Leu Lys Glu Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:37:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                - Pro Val Leu Asp Leu Phe Arg Glu Trp Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:38:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Trp Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:39:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Leu Lys Ala         #                15                                                           - Leu Lys Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:40:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                - Pro Val Leu Asp Leu Phe Asn Glu Leu Leu Ar - #g Glu Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:41:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                - Pro Val Leu Asp Leu Trp Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:42:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Trp Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:43:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu Tr - #p Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:44:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All genetically encoded amino acids                                          #the D-configurationn                                                                   (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:45:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:46:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                - Pro Val Leu Asp Leu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:47:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                - Val Leu Asp Leu Phe Arg Glu Leu Leu Asn Gl - #u Gly Leu Glu Ala Leu         #                15                                                           - Lys Gln Lys Leu Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:48:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = D-Pro) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 2                                                     #Xaa = D-Val) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                - Xaa Xaa Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:49:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                - Pro Val Leu Asp Leu Phe Arg Asn Leu Leu Gl - #u Lys Leu Leu Glu Ala         #                15                                                           - Leu Glu Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:50:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Tr - #p Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:51:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                - Pro Val Leu Asp Leu Phe Trp Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:52:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                - Pro Val Trp Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:53:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                - Val Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:54:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Trp Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:55:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                - Pro Leu Phe Arg Glu Leu Leu Asn Glu Leu Le - #u Glu Ala Leu Lys Gln         #                15                                                           - Lys Leu Lys                                                                 - (2) INFORMATION FOR SEQ ID NO:56:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Lys Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:57:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                - Pro Val Leu Asp Leu Phe Arg Asn Leu Leu Gl - #u Glu Leu Leu Lys Ala         #                15                                                           - Leu Glu Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:58:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:59:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                - Leu Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:60:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln                                                                 - (2) INFORMATION FOR SEQ ID NO:61:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                - Pro Val Leu Asp Glu Phe Arg Trp Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:62:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                - Pro Val Leu Asp Glu Trp Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:63:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                - Pro Val Leu Asp Phe Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:64:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                - Pro Trp Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:65:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 12                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                - Val Leu Asp Glu Phe Arg Glu Lys Leu Asn Gl - #u Xaa Leu Glu Ala Leu         #                15                                                           - Lys Gln Lys Leu Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:66:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                - Pro Val Leu Asp Leu Phe Arg Asn Leu Leu Gl - #u Glu Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:67:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                - Val Leu Asp Leu Phe Arg Glu Leu Leu Asn Gl - #u Leu Leu Glu Ala Leu         #                15                                                           - Lys Gln Lys Leu Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:68:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu Ly - #s Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:69:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                - Pro Val Leu Asp Glu Phe Arg Lys Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:70:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu Ty - #r Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:71:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 14                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Leu Xaa Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:72:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Trp Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:73:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                - Pro Val Leu Asp Glu Phe Trp Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:74:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                - Pro Val Leu Asp Lys Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:75:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:76:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu Ph - #e Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:77:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Lys Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:78:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                - Pro Val Leu Asp Glu Phe Arg Asp Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:79:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:80:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                - Pro Val Leu Asp Leu Phe Glu Arg Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:81:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Trp Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:82:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 11                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                - Leu Asp Glu Phe Arg Glu Lys Leu Asn Glu Xa - #a Leu Glu Ala Leu Lys         #                15                                                           - Gln Lys Leu Lys                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:83:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Trp Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:84:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:85:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                - Pro Leu Asp Leu Phe Arg Glu Leu Leu Asn Gl - #u Leu Leu Glu Ala Leu         #                15                                                           - Lys Gln Lys Leu Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:86:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                - Pro Val Leu Glu Leu Phe Glu Arg Leu Leu As - #p Glu Leu Leu Asn Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:87:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All amino acids are in theTION:                                                             D-configurat - #ion                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                - Pro Leu Leu Glu Leu Leu Lys Glu Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:88:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                - Pro Val Leu Asp Lys Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:89:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Trp Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:90:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 10                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                - Asp Glu Phe Arg Glu Lys Leu Asn Glu Xaa Le - #u Glu Ala Leu Lys Gln         #                15                                                           - Lys Leu Lys                                                                 - (2) INFORMATION FOR SEQ ID NO:91:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:92:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                - Pro Val Leu Asp Glu Phe Arg Glu Leu Tyr As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:93:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Lys Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:94:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Ala Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:95:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Leu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:96:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All genetically encoded amino acids                                          #the D-configurationn                                                                   (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:97:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu             #                15                                                           - 2) INFORMATION FOR SEQ ID NO:98:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:99:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                - Lys Leu Lys Gln Lys Leu Ala Glu Leu Leu Gl - #u Asn Leu Leu Glu Arg         #                15                                                           - Phe Leu Asp Leu Val Pro                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:100:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All amino acids are in theTION:                                                             D-configurat - #ion                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:101:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Trp Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:102:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Leu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Glu Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:103:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                               - Pro Val Leu Asp Glu Phe Arg Glu Leu Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:104:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                               - Pro Leu Leu Asn Glu Leu Leu Glu Ala Leu Ly - #s Gln Lys Leu Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:105:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                               - Pro Ala Ala Asp Ala Phe Arg Glu Ala Ala As - #n Glu Ala Ala Glu Ala         #                15                                                           - Ala Lys Gln Lys Ala Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:106:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                               - Pro Val Leu Asp Leu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:107:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All amino acids are in theTION:                                                             D-configurat - #ion                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                               - Lys Leu Lys Gln Lys Leu Ala Glu Leu Leu Gl - #u Asn Leu Leu Glu Arg         #                15                                                           - Phe Leu Asp Leu Val Pro                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:108:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                               - Pro Val Leu Asp Leu Phe Arg Trp Leu Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:109:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Arg Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:110:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 14                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Xaa Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:111:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu Tr - #p Glu Xaa Trp Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:112:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Ser Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:113:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Pro Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:114:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Met Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:115:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                               - Pro Lys Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:116:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                               - Pro His Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:117:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                               - Pro Glu Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:118:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 13                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Xaa Leu Glu Ala         #                15                                                           - Leu Glu Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:119:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 17                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Xaa Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:120:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 16                                                    #Xaa = Aib(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Xaa         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:121:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Ala         #                15                                                           - Leu Trp Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:122:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                               - Pro Val Leu Asp Glu Phe Arg Glu Lys Leu As - #n Glu Glu Leu Glu Trp         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:123:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                               - Gln Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:124:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 18                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 20                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 22                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                               - Pro Val Leu Asp Leu Phe Xaa Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Xaa Gln Xaa Leu Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:125:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                               - Asn Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:126:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Gly Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:127:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Leu         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:128:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Phe         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:129:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                               - Pro Val Leu Glu Leu Phe Asn Asp Leu Leu Ar - #g Glu Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:130:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                               - Pro Val Leu Glu Leu Phe Asn Asp Leu Leu Ar - #g Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:131:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                               - Pro Val Leu Glu Leu Phe Lys Glu Leu Leu As - #n Glu Leu Leu Asp Ala         #                15                                                           - Leu Arg Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:132:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Asn Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:133:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                               - Pro Val Leu Glu Leu Phe Glu Arg Leu Leu Gl - #u Asp Leu Leu Gln Ala         #                15                                                           - Leu Asn Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:134:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                               - Pro Val Leu Glu Leu Phe Glu Arg Leu Leu Gl - #u Asp Leu Leu Lys Ala         #                15                                                           - Leu Asn Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:135:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                               - Asp Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:136:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                               - Pro Ala Leu Glu Leu Phe Lys Asp Leu Leu Gl - #n Glu Leu Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:137:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 17                                                    #Xaa = NaphthylalanineFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Xaa Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:138:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Trp         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:139:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Trp As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:140:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 18                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 20                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 22                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Xaa Gln Xaa Leu Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:141:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                               - Pro Val Leu Asp Phe Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:142:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                               - Pro Val Leu Glu Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:143:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #N-terminal acetylated andATION:                                              #amidated peptideterminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Gly Leu Glu Ala         #                15                                                           - Leu Lys Gln Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:144:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #Xaa = D-Pro) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                               - Xaa Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:145:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                               - Gly Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:146:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:147:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Phe Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:148:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Gly Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:149:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Trp Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:150:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                               - Pro Leu Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:151:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Gly Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:152:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                               - Pro Val Phe Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:153:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                               - Ala Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:154:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Gly Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:155:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Trp Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:156:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:157:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                               - Pro Val Leu Glu Phe Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:158:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Arg Leu Leu Asp Trp         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:159:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                               - Pro Val Leu Asp Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:160:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Trp         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:161:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:162:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                               - Pro Val Leu Glu Leu Phe Glu Asn Trp Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:163:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Trp Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:164:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Trp Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:165:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Leu         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:166:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:167:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                               - Pro Val Leu Glu Leu Phe Glu Asn Gly Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:168:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                               - Pro Val Leu Glu Leu Phe Glu Gln Leu Leu Gl - #u Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:169:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:170:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 12                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 19                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 20                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 22                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Xaa Leu Leu Asp Ala         #                15                                                           - Leu Gln Xaa Xaa Leu Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:171:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Lys Leu Leu Asp Leu         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:172:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Arg Leu Gly Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:173:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                               - Pro Val Leu Asp Leu Phe Asp Asn Leu Leu As - #p Arg Leu Leu Asp Leu         #                15                                                           - Leu Asn Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:174:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...22                                                #All amino acids are in theTION:                                                             D-configurat - #ion                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:175:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Glu Leu         #                15                                                           - Leu Asn Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:176:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                               - Pro Val Leu Glu Leu Trp Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:177:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                               - Gly Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:178:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                               - Pro Val Leu Glu Leu Phe Asp Asn Leu Leu Gl - #u Lys Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Arg                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:179:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                               - Pro Val Leu Glu Leu Phe Asp Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:180:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                               - Pro Val Leu Glu Leu Phe Asp Asn Leu Leu As - #p Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Arg                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:181:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Trp Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:182:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Lys Leu Leu Glu Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:183:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                               - Pro Leu Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:184:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                               - Pro Val Leu Glu Leu Phe Leu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Trp Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:185:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 19                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 20                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 22                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Xaa Xaa Leu Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:186:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                               - Pro Val Leu Glu Leu Phe Glu Gln Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:187:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Asn Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:188:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                               - Pro Val Leu Glu Leu Phe Glu Asn Leu Leu As - #p Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:189:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:                               - Asp Val Leu Glu Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Lys                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:190:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:                               - Pro Val Leu Glu Phe Trp Asp Asn Leu Leu As - #p Lys Leu Leu Asp Ala         #                15                                                           - Leu Gln Lys Lys Leu Arg                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:191:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:191:                               - Pro Val Leu Asp Leu Leu Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:192:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192:                               - Pro Val Leu Asp Leu Phe Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:193:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:193:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:194:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:194:                               - Pro Val Leu Glu Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:195:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:195:                               - Pro Val Leu Glu Leu Phe Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:196:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:196:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Asn Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:197:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:197:                               - Pro Leu Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:198:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:198:                               - Gly Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:199:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:199:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Trp Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:200:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:200:                               - Asn Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:201:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:201:                               - Pro Leu Leu Asp Leu Phe Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:202:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:202:                               - Pro Ala Leu Glu Leu Phe Lys Asp Leu Leu Gl - #u Glu Leu Arg Gln Lys         #                15                                                           - Leu Arg                                                                     - (2) INFORMATION FOR SEQ ID NO:203:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:203:                               - Ala Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:204:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:204:                               - Pro Val Leu Asp Phe Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:205:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:205:                               - Pro Val Leu Asp Leu Phe Arg Glu Trp Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:206:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:206:                               - Pro Leu Leu Glu Leu Leu Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:207:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:207:                               - Pro Val Leu Glu Leu Leu Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:208:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:208:                               - Pro Ala Leu Glu Leu Phe Lys Asp Leu Leu Gl - #u Glu Leu Arg Gln Arg         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:209:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:209:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:210:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:210:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:211:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                               (A) NAME/KEY: Other                                                           (B) LOCATION: 14                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 16                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 18                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:211:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Xaa Gln Xaa         #                15                                                           - Leu Xaa                                                                     - (2) INFORMATION FOR SEQ ID NO:212:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                               (A) NAME/KEY: Other                                                           (B) LOCATION: 7                                                     #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 14                                                    #Xaa = Orn(D) OTHER INFORMATION:                                                        (A) NAME/KEY: Other                                                           (B) LOCATION: 16                                                    #Xaa = Orn(D) OTHER INFORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:212:                               - Pro Val Leu Asp Leu Phe Xaa Glu Leu Leu Gl - #u Glu Leu Xaa Gln Xaa         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:213:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:213:                               - Pro Ala Leu Glu Leu Phe Lys Asp Leu Leu Gl - #u Glu Phe Arg Gln Arg         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:214:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                               (A) NAME/KEY: Other                                                           (B) LOCATION: 1                                                     #D-configuration of ProORMATION:                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:214:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:215:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:215:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Trp Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:216:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:216:                               - Pro Val Leu Glu Leu Phe Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:217:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:217:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Leu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:218:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:218:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Leu Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:219:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:219:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Trp Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:220:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:220:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Gln Lys Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:221:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:221:                               - Asp Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:222:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:222:                               - Pro Val Leu Asp Ala Phe Arg Glu Leu Leu Gl - #u Ala Leu Leu Gln Leu         #                15                                                           - Lys Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:223:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:223:                               - Pro Val Leu Asp Ala Phe Arg Glu Leu Leu Gl - #u Ala Leu Ala Gln Leu         #                15                                                           - Lys Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:224:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:224:                               - Pro Val Leu Asp Leu Phe Arg Glu Gly Trp Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:225:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:225:                               - Pro Val Leu Asp Ala Phe Arg Glu Leu Ala Gl - #u Ala Leu Ala Gln Leu         #                15                                                           - Lys Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:226:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:226:                               - Pro Val Leu Asp Ala Phe Arg Glu Leu Gly Gl - #u Ala Leu Leu Gln Leu         #                15                                                           - Lys Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:227:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:227:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Gly Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:228:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:228:                               - Pro Val Leu Asp Leu Phe Arg Glu Gly Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:229:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:229:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Gly Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:230:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:230:                               - Pro Val Leu Glu Leu Phe Glu Arg Leu Leu Gl - #u Asp Leu Gln Lys Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:231:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:231:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Lys Leu Glu Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:232:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:232:                               - Pro Leu Leu Glu Leu Phe Lys Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - Leu Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:233:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:233:                               - This sequence has been intentionally skipped                                - (2) INFORMATION FOR SEQ ID NO:234:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:234:                               - This sequence has been intentionally skipped                                - (2) INFORMATION FOR SEQ ID NO:235:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          -    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:235:                                -  This sequence has been intentionally skipped                               - (2) INFORMATION FOR SEQ ID NO:236:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:236:                               - This sequence has been intentionally skipped                                - (2) INFORMATION FOR SEQ ID NO:237:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:237:                               - Leu Asp Asp Leu Leu Gln Lys Trp Ala Glu Al - #a Phe Asn Gln Leu Leu         #                15                                                           - Lys Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:238:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:238:                               - Glu Trp Leu Lys Ala Phe Tyr Glu Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:239:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:239:                               - Glu Trp Leu Glu Ala Phe Tyr Lys Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:240:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:240:                               - Asp Trp Leu Lys Ala Phe Tyr Asp Lys Val Al - #a Glu Lys Leu Lys Glu         #                15                                                           - Ala Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:241:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:241:                               - Asp Trp Phe Lys Ala Phe Tyr Asp Lys Val Ph - #e Glu Lys Phe Lys Glu         #                15                                                           - Phe Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:242:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:242:                               - Gly Ile Lys Lys Phe Leu Gly Ser Ile Trp Ly - #s Phe Ile Lys Ala Phe         #                15                                                           - Val Gly                                                                     - (2) INFORMATION FOR SEQ ID NO:243:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:243:                               - Asp Trp Phe Lys Ala Phe Tyr Asp Lys Val Al - #a Glu Lys Phe Lys Glu         #                15                                                           - Ala Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:244:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:244:                               - Asp Trp Leu Lys Ala Phe Tyr Asp Lys Val Al - #a Glu Lys Leu Lys Glu         #                15                                                           - Ala Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:245:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:245:                               - Asp Trp Leu Lys Ala Phe Tyr Asp Lys Val Ph - #e Glu Lys Phe Lys Glu         #                15                                                           - Phe Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:246:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:246:                               - Glu Trp Leu Glu Ala Phe Tyr Lys Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:247:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:247:                               - Asp Trp Phe Lys Ala Phe Tyr Asp Lys Phe Ph - #e Glu Lys Phe Lys Glu         #                15                                                           - Phe Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:248:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:248:                               - Glu Trp Leu Lys Ala Phe Tyr Glu Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:249:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:249:                               - Glu Trp Leu Lys Ala Glu Tyr Glu Lys Val Gl - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:250:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:250:                               - Glu Trp Leu Lys Ala Glu Tyr Glu Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:251:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 18 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...18                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:251:                               - Glu Trp Leu Lys Ala Phe Tyr Lys Lys Val Le - #u Glu Lys Leu Lys Glu         #                15                                                           - Leu Phe                                                                     - (2) INFORMATION FOR SEQ ID NO:252:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...15                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:252:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Gln Lys Leu Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:253:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...16                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:253:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Glu Leu Lys Gln Lys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:254:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...16                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:254:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Lys Leu Lys Gln Lys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:255:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...15                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:255:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Lys Leu Gln Lys             #                15                                                           - (2) INFORMATION FOR SEQ ID NO:256:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...16                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:256:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu Gl - #u Ala Leu Lys Gln Lys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:257:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...16                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:257:                               - Pro Val Leu Asp Leu Phe Glu Asn Leu Leu Gl - #u Arg Leu Lys Gln Lys         #                15                                                           - (2) INFORMATION FOR SEQ ID NO:258:                                          -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 16 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: None                                                -     (ix) FEATURE:                                                                     (A) NAME/KEY: Other                                                           (B) LOCATION: 1...16                                                #N-terminal acetylated andATION:                                              #amidated      C-terminal                                                     -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:258:                               - Pro Val Leu Asp Leu Phe Arg Glu Leu Leu As - #n Glu Leu Lys Gln Lys         #                15                                                           __________________________________________________________________________

What is claimed is:
 1. An ApoA-I agonist compound comprising:(i) an 18to 22-residue peptide or peptide analogue which forms an amphipathicα-helix in the presence of lipids and which comprises formula (I):

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p); X₂ is analiphatic amino acid; X₃ is Leu (L); X₄ is an acidic amino acid; X₅ isLeu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basic amino acid;X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ is Leu (L) orTrp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is an acidic aminoacid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basic amino acid orLeu (L); X₁₅ is Gln (Q) or Asn (N); X₁₆ is a basic amino acid; X₁₇ isLeu (L); X₁₈ is a basic amino acid; Z₁ is H₂ N-- or RC(O)NH--; Z₂ is--C(O)NRR, --C(O)OR or --C(O)OH or a salt thereof; each R isindependently --H, (C₁ -C₆) alkyl, (C₁ -C₆) alkenyl, (C₁ -C₆) alkynyl,(C₅ -C₂₀) aryl, (C₆ -C₂₆) alkaryl, 5-20 membered heteroaryl or 6-26membered alkheteroaryl or a 1 to 4-residue peptide or peptide analoguein which one or more bonds between residues 1-7 are independently asubstituted amide, an isostere of an amide or an amide mimetic; and each"-" between residues X₁ through X₁₈ independently designates an amidelinkage, a substituted amide linkage, an isostere of an amide or anamide mimetic; or (ii) an altered form of formula (I) in which at leastone of residues X₁, X₂, X₃, X₄, X₅, X₆, X₇, X₈, X₉, X₁₀, X₁₁, X₁₂, X₁₃,X₁₄, X₁₅, X₁₆, X₁₇ or X₁₈ is conservatively substituted with anotherresidue.
 2. The ApoA-I agonist compound of claim 1 which exhibits atleast about 38% LCAT-activation activity as compared with human ApoA-I.3. The ApoA-I agonist compound of claim 1 which is the altered form offormula (I).
 4. The ApoA-I agonist compound of claim 3 in which residuesX₂, X₃, X₅, X₆, X₉, X₁₀, X₁₃ and X₁₇ are fixed according to formula (I)and at least one of residues X₁, X₄, X₇, X₈, X₁₁, X₁₂, X₁₄, X₁₅, X₁₆ andX₁₈ is conservatively substituted with another residue.
 5. The ApoA-Iagonist compound of claim 4 in which:X₁ is Pro (P), D-Pro (p), Gly (G),Asn (N) or Ala (A); X₂ is Ala (A), Leu (L) or Val (V); X₃ is Leu (L); X₅is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₉ is Leu (L) or Trp(W); X₁₀ is Leu (L) or Trp (W); X₁₃ is Leu (L), Trp (W) or Phe (F); andX₁₇ is Leu (L).
 6. The ApoA-I agonist compound of claim 3 in which X₄,X₇, X₈, X₁₁, X₁₂, X₁₄, X₁₅ and X₁₈ are fixed according to formula (I)and at least one of residues X₁, X₂, X₃, X₅, X₆, X₉, X₁₀, X₁₃, X₁₆ andX₁₇ is conservatively substituted with another residue.
 7. The ApoA-Iagonist compound of claim 6 in which:X₄ is Asp (D) or Glu (E); X₇ is Arg(R), Lys (K) or Orn; X₈ is Asp (D) or Glu (E); X₁₁ is Asn (N) or Glu(E); X₁₂ is Glu (E); X₁₄ is Lys (K), Arg (R) or Orn; X₁₅ is Gln (Q) orAsn (N); X₁₆ is Lys (K), Arg (R) or Orn; and X₁₈ is His (H), Lys (K) orArg (R).
 8. The ApoA-I agonist compound of claim 7 in which X₃ is Leu(L), X₆ is Phe (F), X₉ is Leu (L) or Trp (W), X₁₀ is Leu (L) or Trp (W).9. The ApoA-I agonist compound of claim 5 or 7 in which theconservatively substituting residue is classified within a samesub-category as the conservatively substituted residue.
 10. The ApoA-Iagonist compound of claim 1 which is an 18-residue peptide or peptideanalogue according to formula (I).
 11. The ApoA-I agonist compound ofclaim 10 in which:the "-" between residues X₁ through X₁₈ designates--C(O)NH--; Z₁ is H₂ N--; and Z₂ is --C(O)OH or a salt thereof.
 12. TheApoA-I agonist compound of claim 11, in which:X₁ is Pro (P), Ala (A),Gly (G), Asn (N) or D-Pro (p); X₂ is Ala (A), Val (V) or Leu (L); X₃ isLeu (L); X₄ is Asp (D) or Glu (E); X₅ is Leu (L) or Phe (F); X₆ is Leu(L) or Phe (F); X₇ is Arg (R), Lys (K) or Orn; X₈ is Asp (D) or Glu (E);X₉ is Leu (L) or Trp (W); X₁₀ is Leu (L) or Trp (W); X₁₁ is Glu (E) orAsn (N); X₁₂ is Glu (E); X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is Arg(R), Lys (K) or Orn; X₁₅ is Gln (Q) or Asn (N); X₁₆ is Arg (R), Lys (K)or Orn; X₁₇ is Leu (L); and X₁₈ is Arg (R), Lys (K) or Orn.
 13. TheApoA-I agonist compound of claim 1 which is selected from the groupconsisting of:peptide 191 PVLDLLRELLEELKQKLK* (SEQ ID NO:191); peptide192 PVLDLFKELLEELKQKLK* (SEQ ID NO:192); peptide 193 PVLDLFRELLEELKQKLK*(SEQ ID NO:193); peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194);peptide 195 PVLELFKELLEELKQKLK* (SEQ ID NO:195); peptide 196PVLDLFRELLEELKNKLK* (SEQ ID NO:196); peptide 197 PLLDLFRELLEELKQKLK*(SEQ ID NO:197); peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198);peptide 199 PVLDLFRELWEELKQKLK* (SEQ ID NO:199); peptide 200NVLDLFRELLEELKQKLK* (SEQ ID NO:200); peptide 201 PLLDLFKELLEELKQKLK*(SEQ ID NO:201); peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202);peptide 203 AVLDLFRELLEELKQKLK* (SEQ ID NO:203); peptide 204PVLDFFRELLEELKQKLK* (SEQ ID NO:204); peptide 205 PVLDLFREWLEELKQKLK*(SEQ ID NO:205); peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206);peptide 207 PVLELLKELLEELKQKLK* (SEQ ID NO:207); peptide 208PALELFKDLLEELRQRLK* (SEQ ID NO:208); peptide 209 PVLDLFRELLNELLQKLK (SEQID NO:209); peptide 210 PVLDLFRELLEELKQKLK (SEQ ID NO:210); peptide 211PVLDLFRELLEELOQOLO* (SEQ ID NO:211); peptide 212 PVLDLFOELLEELOQOLK*(SEQ ID NO:212); peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213);peptide 214 pVLDLFRELLEELKQKLK* (SEQ ID NO:214); peptide 215PVLDLFRELLEEWKQKLK* (SEQ ID NO:215); peptide 230 PVLELFERLLEDLQKKLK (SEQID NO:230); peptide 231 PVLDLFRELLEKLEQKLK (SEQ ID NO:231); and peptide232 PLLELFKELLEELKQKLK* (SEQ ID NO:232);in either N- and/or C-terminalblocked or unblocked forms.
 14. A multimeric ApoA-I agonist compoundwhich exhibits at least about 38% LCAT activation activity as comparedwith human ApoA-I and which has the structural formula (II): ##STR6## ora pharmaceutically acceptable salt thereof, wherein: each m isindependently an integer from 0 to 1;n is an integer from 0 to 10; each"HH" is independently a compound according to claim 1; each "LL" isindependently a bifunctional linker; and each "-" independentlydesignates a covalent linkage.
 15. A multimeric ApoA-I agonist compoundwhich exhibits at least about 38% LCAT activation activity as comparedwith human ApoA-I and which has the structural formula (III): ##STR7##or a pharmaceutically acceptable salt thereof, wherein: each X isindependently ##STR8## each HH is independently a compound according toclaim 1; each LL is independently a bifunctional linker;each m isindependently an integer from 0 to 1; each n is independently an integerfrom 0 to 8; N_(ya) and N_(yb) are each independently a multifunctionallinking moiety where y_(a) and y_(b) represent the number of functionalgroups on N_(ya) and N_(yb), respectively; each y_(a) or y_(b) isindependently an integer from 3 to 8; p is an integer from 0 to 7; andeach "-" independently designates a covalent bond.
 16. A multimericApoA-I compound agonist which exhibits at least about 38% LCATactivation activity as compared with human ApoA-I and which has thestructural formula (IV) or (V): ##STR9## or a pharmaceuticallyacceptable salt thereof, wherein: each X is independently ##STR10## eachHH is independently a compound according to claim 1; each LL isindependently a bifunctional linker;each n is independently an integerfrom 0 to 1; each m is independently an integer from 0 to 8; R₁ is --ORor --NRR; and each R is independently --H, (C₁ -C₆) alkyl, (C₁ -C₆)alkenyl, (C₁ -C₆) alkynyl, (C₅ -C₂₀) aryl, (C₆ -C₂₆) alkaryl, 5-20membered heteroaryl or 6-26 membered alkheteroaryl.
 17. The multimericApoA-I agonist compound of claim 14, 15 or 16 in which the bifunctionallinker is cleavable.
 18. The ApoA-I multimeric agonist compound of claim14, 15 or 16 in which n is
 0. 19. The multimeric ApoA-I agonist compoundof claim 18 in which m is
 0. 20. The multimeric ApoA-I agonist compoundof claim 14, 15 or 16 in which each HH is independently an 18-residuepeptide or peptide analogue of formula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);X₂ is an aliphatic amino acid; X₃ is Leu (L); X₄ is an acidic aminoacid; X₅ is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basicamino acid; X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ isLeu (L) or Trp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is anacidic amino acid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basicamino acid or Leu (L); X₁₅ is Gln (Q) or Asn (N); X₁₆ is a basic aminoacid; X₁₇ is Leu (L); X₁₈ is a basic amino acid; each "-" betweenresidues X₁ through X₁₈ designates --C(O)NH--; Z₁ is H₂ N--; and Z₂ is--C(O)OH or a salt thereof.
 21. The multimeric ApoA-I agonist compoundof claim 14, 15 or 16 in which each HH is independently an 18-residuepeptide or peptide analogue of formula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N) or D-Pro (p); X₂ isAla (A), Val (V) or Leu (L); X₃ is Leu (L); X₄ is Asp (D) or Glu (E); X₅is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is Arg (R), Lys (K)or Orn; X₈ is Asp (D) or Glu (E); X₉ is Leu (L) or Trp (W); X₁₀ is Leu(L) or Trp (W); X₁₁ is Glu (E) or Asn (N); X₁₂ is Glu (E); X₁₃ is Leu(L), Trp (W) or Phe (F); X₁₄ is Arg (R), Lys (K) or Orn; X₁₅ is Gln (Q)or Asn (N); X₁₆ is Arg (R), Lys (K) or Orn; X₁₇ is Leu (L); X₁₈ is Arg(R), Lys (K) or Orn; each "-" between residues X₁ through X₁₈ designates--C(O)NH--; Z₁ is H₂ N--; and Z₂ is --C(O)OH or a salt thereof.
 22. Themultimeric ApoA-I agonist compound of claim 14, 15 or 16 in which eachHH is independently a peptide selected from the group consistingof:peptide 191 PVLDLLRELLEELKQKLK* (SEQ ID NO:191); peptide 192PVLDLFKELLEELKQKLK* (SEQ ID NO:192); peptide 193 PVLDLFRELLEELKQKLK*(SEQ ID NO:193); peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194);peptide 195 PVLELFKELLEELKQKLK* (SEQ ID NO:195); peptide 196PVLDLFRELLEELKNKLK* (SEQ ID NO:196); peptide 197 PLLDLFRELLEELKQKLK*(SEQ ID NO:197); peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198);peptide 199 PVLDLFRELWEELKQKLK* (SEQ ID NO:199); peptide 200NVLDLFRELLEELKQKLK* (SEQ ID NO:200); peptide 201 PLLDLFKELLEELKQKLK*(SEQ ID NO:201); peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202);peptide 203 AVLDLFRELLEELKQKLK* (SEQ ID NO:203); peptide 204PVLDFFRELLEELKQKLK* (SEQ ID NO:204); peptide 205 PVLDLFREWLEELKOKLK*(SEQ ID NO:205): peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206);peptide 207 PVLELLKELLEELKQKLK* (SEQ ID NO:207): peptide 208PALELFKDLLEELRQRLK* (SEQ ID NO:208); peptide 209 PVLDLFRELLNELLQKLK (SEQID NO:209); peptide 210 PVLDLFRELLEELKQKLK (SEQ ID NO:210); peptide 211PVLDLFRELLEELOQOLO* (SEQ ID NO:211); peptide 212 PVLDLFOELLEELOQOLK*(SEQ ID NO:212); peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213);peptide 214 pVLDLFRELLEELKQKLK* (SEQ ID NO:214); peptide 215PVLDLFRELLEEWKQKLK* (SEQ ID NO:215); peptide 230 PVLELFERLLEDLQKKLK (SEQID NO:230); peptide 231 PVLDLFRELLEKLEQKLK (SEQ ID NO:231); and peptide232 PLLELFKELLEELKQKLK* (SEQ ID NO:232);in either N- and/or C-terminalblocked or unblocked forms.
 23. An ApoA-I agonist-lipid complexcomprising an ApoA-I agonist and a lipid, wherein the ApoA-I agonist isa peptide or peptide analogue according to claim 1, a multimeric ApoA-Iagonist compound according to claim 14, a multimeric ApoA-I agonistcompound according to claim 15, or a multimeric ApoA-I agonist compoundaccording to claim
 16. 24. The ApoA-I agonist-lipid complex of claim 23in which the ApoA-I agonist compound is a peptide or peptide analogue offormula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);X₂ is an aliphatic amino acid; X₃ is Leu (L); X₄ is an acidic aminoacid; X₅ is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basicamino acid; X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ isLeu (L) or Trp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is anacidic amino acid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basicamino acid or Leu (L); X₁₅ is Gln (Q) or Asn (N); X₁₆ is a basic aminoacid; X₁₇ is Leu (L); X₁₈ is a basic amino acid; Z₁ is H₂ N-- orRC(O)NH--; Z₂ is --C(O)NRR, --C(O)OR or --C(O)OH or a salt thereof; eachR is independently --H, (C₁ -C₆) alkyl, (C₁ -C₆) alkenyl, (C₁ -C₆)alkynyl, (C₅ -C₂₀) aryl, (C₆ -C₂₆) alkaryl, 5-20 membered heteroaryl or6-26 membered alkheteroaryl or a 1 to 4-residue peptide or peptideanalogue in which one or more bonds between residues 1-7 areindependently a substituted amide, an isostere of an amide or an amidemimetic; and each "-" between residues X₁ through X₁₈ independentlydesignates an amide linkage, a substituted amide linkage, an isostere ofan amide or an amide mimetic.
 25. The ApoA-I agonist-lipid complex ofclaim 23 in which the ApoA-I agonist is a peptide or peptide analogue offormula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);X₂ is an aliphatic amino acid; X₃ is Leu (L); X₄ is an acidic aminoacid; X₅ is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basicamino acid; X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ isLeu (L) or Trp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is anacidic amino acid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basicamino acid or Leu (L); X₁₅ is Gln (Q) or Asn (N); X₁₆ is a basic aminoacid; X₁₇ is Leu (L); X₁₈ is a basic amino acid; the "-" betweenresidues designates --C(O)NH--; Z₁ is H₂ N--; and Z₂ is --C(O)OH or asalt thereof.
 26. The ApoA-I agonist-lipid complex of claim 23 in whichthe ApoA-I agonist is a peptide or peptide analogue of formula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N) or D-Pro (p); X₂ isAla (A), Val (V) or Leu (L); X₃ is Leu (L); X₄ is Asp (D) or Glu (E); X₅is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is Arg (R), Lys (K)or Orn; X₈ is Asp (D) or Glu (E); X₉ is Leu (L) or Trp (W); X₁₀ is Leu(L) or Trp (W); X₁₁ is Glu (E) or Asn (N); X₁₂ is Glu (E); X₁₃ is Leu(L), Trp (W) or Phe (F); X₁₄ is Arg (R), Lys (K) or Orn; X₁₅ is Gln (Q)or Asn (N); X₁₆ is Arg (R), Lys (K) or Orn; X₁₇ is Leu (L); and X₁₈ isArg (R), Lys (K) or Orn; the "-" between residues designates --C(O)NH--;Z₁ is H₂ N--; and Z₂ is --C(O)OH or a salt thereof.
 27. The ApoA-Iagonist-lipid complex of claim 23 in which the ApoA-I agonist is apeptide selected from the group consisting of:peptide 191PVLDLLRELLEELKQKLK* (SEQ ID NO:191); peptide 192 PVLDLFKELLEELKQKLK*(SEQ ID NO:192); peptide 193 PVLDLFRELLEELKQKLK* (SEQ ID NO:193);peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194); peptide 195PVLELFKELLEELKQKLK* (SEQ ID NO:195); peptide 196 PVLDLFRELLEELKNKLK*(SEQ ID NO:196); peptide 197 PLLDLFRELLEELKQKLK* (SEQ ID NO:197);peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198); peptide 199PVLDLFRELWEELKQKLK* (SEQ ID NO:199); peptide 200 NVLDLFRELLEELKQKLK*(SEQ ID NO:200); peptide 201 PLLDLFKELLEELKQKLK* (SEQ ID NO:201);peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202); peptide 203AVLDLFRELLEELKQKLK* (SEQ ID NO:203); peptide 204 PVLDFFRELLEELKQKLK*(SEQ ID NO:204); peptide 205 PVLDLFREWLEELKOKLK* (SEQ ID NO:205);peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206); peptide 207PVLELLKELLEELKQKLK* (SEQ ID NO:207); peptide 208 PALELFKDLLEELRQRLK*(SEQ ID NO:208); peptide 209 PVLDLFRELLNELLQKLK (SEQ ID NO:209); peptide210 PVLDLFRELLEELKQKLK (SEQ ID NO:210); peptide 211 PVLDLFRELLEELOQOLO*(SEQ ID NO:211); peptide 212 PVLDLFOELLEELOQOLK* (SEQ ID NO:212);peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213); peptide 214pVLDLFRELLEELKQKLK* (SEQ ID NO:214); peptide 215 PVLDLFRELLEEWKQKLK*(SEQ ID NO:215); peptide 230 PVLELFERLLEDLQKKLK (SEQ ID NO:230); peptide231 PVLDLFRELLEKLEQKLK (SEQ ID NO:231); and peptide 232PLLELFKELLEELKQKLK* (SEQ ID NO:232);in either N- and/or C-terminalblocked or unblocked forms.
 28. The ApoA-I agonist-lipid complex ofclaim 23 in which the lipid is sphingomyelin.
 29. The ApoA-Iagonist-lipid complex of claim 23 which is in the form of a lyophilizedpowder.
 30. The ApoA-I agonist-lipid complex of claim 23 which is in theform of a solution.
 31. A pharmaceutical composition comprising anApoA-I agonist and a pharmaceutically acceptable carrier, excipient ordiluent, wherein the ApoA-I agonist is a compound according to claim 1,a multimeric ApoA-I agonist compound according to claim 14, a multimericApoA-I agonist compound according to claim 15, or a multimeric ApoA-Iagonist compound according to claim
 16. 32. The pharmaceuticalcomposition of claim 31 in which the ApoA-I agonist compound is apeptide or peptide analogue of formula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);X₂ is an aliphatic amino acid; X₃ is Leu (L); X₄ is an acidic aminoacid; X₅ is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basicamino acid; X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ isLeu (L) or Trp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is anacidic amino acid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basicamino acid or Leu (L); X₁₅ is Gin (Q) or Asn (N); X₁₆ is a basic aminoacid; X₁₇ is Leu (L); X₁₈ is a basic amino acid; Z₁ is H₂ N-- orRC(O)NH--; Z₂ is --C(O)NRR, --C(O)OR or --C(O)OH or a salt thereof; eachR is independently --H, (C₁ -C₆) alkyl, (C₁ -C₆) alkenyl, (C₁ -C₆)alkynyl, (C₅ -C₂₀) aryl, (C₆ -C₂₆) alkaryl, 5-20 membered heteroaryl or6-26 membered alkheteroaryl or a 1 to 4-residue peptide or peptideanalogue in which one or more bonds between residues 1-7 areindependently a substituted amide, an isostere of an amide or an amidemimetic; and each "-" between residues X₁ through X₁₈ independentlydesignates an amide linkage, a substituted amide linkage, an isostere ofan amide or an amide mimetic.
 33. The pharmaceutical composition ofclaim 31 in which the ApoA-I agonist compound is a peptide or peptideanalogue of formula (I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N), Gln (Q) or D-Pro (p);X₂ is an aliphatic amino acid; X₃ is Leu (L); X₄ is an acidic aminoacid; X₅ is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is a basicamino acid; X₈ is an acidic amino acid; X₉ is Leu (L) or Trp (W); X₁₀ isLeu (L) or Trp (W); X₁₁ is an acidic amino acid or Asn (N); X₁₂ is anacidic amino acid; X₁₃ is Leu (L), Trp (W) or Phe (F); X₁₄ is a basicamino acid or Leu (L); X₁₅ is Gln (Q) or Asn (N); X₁₆ is a basic aminoacid; X₁₇ is Leu (L); X₁₈ is a basic amino acid; the "-" betweenresidues designates --C(O)NH--; Z₁ is H₂ N--; and Z₂ is --C(O)OH or asalt thereof.
 34. The pharmaceutical composition of claim 31 in whichthe ApoA-I agonist compound is a peptide or peptide analogue of formula(I)

    Z.sub.1 -X.sub.1 -X.sub.2 -X.sub.3 -X.sub.4 -X.sub.5 -X.sub.6 -X.sub.7 -X.sub.8 -X.sub.9 -X.sub.10 -X.sub.11 -X.sub.12 -X.sub.13 -X.sub.14 -X.sub.15 -X.sub.16 -X.sub.17 -X.sub.18 -Z.sub.2          (I)

wherein: X₁ is Pro (P), Ala (A), Gly (G), Asn (N) or D-Pro (p); X₂ isAla (A), Val (V) or Leu (L); X₃ is Leu (L); X₄ is Asp (D) or Glu (E); X₅is Leu (L) or Phe (F); X₆ is Leu (L) or Phe (F); X₇ is Arg (R), Lys (K)or Orn; X₈ is Asp (D) or Glu (E); X₉ is Leu (L) or Trp (W); X₁₀ is Leu(L) or Trp (W); X₁₁ is Glu (E) or Asn (N); X₁₂ is Glu (E); X₁₃ is Leu(L), Trp (W) or Phe (F); X₁₄ is Arg (R), Lys (K) or Orn; X₁₅ is Gln (Q)or Asn (N); X₁₆ is Arg (R), Lys (K) or Orn; X₁₇ is Leu (L); and X₁₈ isArg (R), Lys (K) or Orn; the "-" between residues designates --C(O)NH--;Z₁ is H₂ N--; and Z₂ is --C(O)OH or a salt thereof.
 35. Thepharmaceutical composition of claim 31 in which the ApoA-I agonistcompound is a peptide selected from the group consisting of:peptide 191PVLDLLRELLEELKQKLK* (SEQ ID NO:191); peptide 192 PVLDLFKELLEELKQKLK*(SEQ ID NO:192); peptide 193 PVLDLFRELLEELKQKLK* (SEQ ID NO:193);peptide 194 PVLELFRELLEELKQKLK* (SEQ ID NO:194); peptide 195PVLELFKELLEELKQKLK* (SEQ ID NO:195); peptide 196 PVLDLFRELLEELKNKLK*(SEQ ID NO:196); peptide 197 PLLDLFRELLEELKQKLK* (SEQ ID NO:197);peptide 198 GVLDLFRELLEELKQKLK* (SEQ ID NO:198); peptide 199PVLDLFRELWEELKQKLK* (SEQ ID NO:199); peptide 200 NVLDLFRELLEELKQKLK*(SEQ ID NO:200); peptide 201 PLLDLFKELLEELKQKLK* (SEQ ID NO:201);peptide 202 PALELFKDLLEELRQKLR* (SEQ ID NO:202); peptide 203AVLDLFRELLEELKQKLK* (SEQ ID NO:203); peptide 204 PVLDFFRELLEELKQKLK*(SEQ ID NO:204); peptide 205 PVLDLFREWLEELKQKLK* (SEQ ID NO:205);peptide 206 PLLELLKELLEELKQKLK* (SEQ ID NO:206); peptide 207PVLELLKELLEELKQKLK* (SEQ ID NO:207); peptide 208 PALELFKDLLEELRQRLK*(SEQ ID NO:208); peptide 209 PVLDLFRELLNELLQKLK (SEQ ID NO:209); peptide210 PVLDLFRELLEELKQKLK (SEQ ID NO:210); peptide 211 PVLDLFRELLEELOQOLO*(SEQ ID NO:211); peptide 212 PVLDLFOELLEELOQOLK* (SEQ ID NO:212);peptide 213 PALELFKDLLEEFRQRLK* (SEQ ID NO:213); peptide 214pVLDLFRELLEELKQKLK* (SEQ ID NO:214); peptide 215 PVLDLFRELLEEWKQKLK*(SEQ ID NO:215); peptide 230 PVLELFERLLEDLQKKLK (SEQ ID NO:230); peptide231 PVLDLFRELLEKLEQKLK (SEQ ID NO:231); and peptide 232PLLELFKELLEELKQKLK* (SEQ ID NO:232);in either N- and/or C-terminalblocked or unblocked forms.
 36. The pharmaceutical composition of claim31 in which the ApoA-I agonist is in the form of an ApoA-I agonist-lipidcomplex, said complex comprising the ApoA-I agonist compound and alipid.
 37. The pharmaceutical composition of claim 36 in which theApoA-I agonist-lipid complex is in the form of a lyophilized powder. 38.A method of treating a subject suffering from a disorder associated withdyslipidemia, said method comprising the step of administering to thesubject an effective amount of the ApoA-I agonist compound of claim 1.39. The method of claim 38 in which the ApoA-I agonist compound is inthe form of a pharmaceutical composition, said composition comprisingthe ApoA-I agonist compound and a pharmaceutically acceptable carrier,excipient or diluent.
 40. The method of claim 38 in which the ApoA-Iagonist compound is in the form of an ApoA-I agonist-lipid complex, saidcomplex comprising the ApoA-I agonist compound and a lipid.
 41. Themethod of claim 38 in which the disorder associated with dyslipidemia ishypercholesterolemia.
 42. The method of claim 38 in which the disorderassociated with dyslipidemia is cardiovascular disease.
 43. The methodof claim 38 in which the disorder associated with dyslipidemia isatherosclerosis.
 44. The method of claim 38 in which the disorderassociated with dyslipidemia is restenosis.
 45. The method of claim 38,in which the disorder associated with dyslipidemia is HDL or ApoA-Ideficiency.
 46. The method of claim 38, in which the disorder associatedwith dyslipidemia is hypertriglyceridemia.
 47. The method of claim 38,in which the disorder associated with dyslipidemia is metabolicsyndrome.
 48. A method of treating a subject suffering from septicshock, said method comprising the step of administering to the subjectan effective amount of the ApoA-I agonist compound of claim
 1. 49. Themethod of claim 38 or 48 in which said subject is a human.
 50. Themethod of claim 38 or 48 in which about 0.5 mg/kg to about 100 mg/kgApoA-I agonist compound is administered to said subject.
 51. Thepharmaceutical composition of claim 32 in which the ApoA-I agonist is inthe form of an ApoA-I agonist-lipid complex, said complex comprising theApoA-I agonist compound and a lipid.
 52. The pharmaceutical compositionof claim 33 in which the ApoA-I agonist is in the form of an ApoA-Iagonist-lipid complex, said complex comprising the ApoA-I agonistcompound and a lipid.
 53. The pharmaceutical composition of claim 34 inwhich the ApoA-I agonist is in the form of an ApoA-I agonist-lipidcomplex, said complex comprising the ApoA-I agonist compound and alipid.
 54. The pharmaceutical composition of claim 35 in which theApoA-I agonist is in the form of an ApoA-I agonist-lipid complex, saidcomplex comprising the ApoA-I agonist compound and a lipid.